,transcrib]'s notes3 ,ap>t f ! foll[+ correct$ misspell+s ! text ( ?is book has be5 pres]v$ z 9 ! orig9al3 xolol -- xylol side -- slide ov]/ra9+ -- ov]/a9+ ,:]e appropriate1 illu/rations & footnotes h be5 position$ adjac5t to ! relevant text4 ,,section ,,cutt+ ,,& ,,/a9+ ,a ,,practical ,,introduction ,,to ,,hi/ological ,,me?ods ,,= ,,/ud5ts ,,& ,,practition]s ,,by ,w4 ,s4 ,,colman1 ,m4,d41 ,m4,r4,c4,p4 ,,assi/ant ,,physician 7,,=m]ly ,,pa?ologi/7 ,,to ,,! ,,national ,,hospital ,,= ,,! ,,p>alys$ ,,& ,,epileptic2 ,,& ,,to ,,! ,,hospital ,,= ,,sick ,,*ildr51 ,,great ,,ormond ,,/reet1 ,,etc4 ,,second ,,edition ,,enl>g$ ,,& ,,9 ,,mo/ ,,"p ,,re-,,writt5 ,,london ,h4 ,k4 ,,lewis1 #acf ,,g[] ,,/reet1 ,w4,c4 #ahif ,,pr9t$ ,,by ,h4 ,k4 ,,lewis1 #acf ,,g[] ,,/reet1 ,,london1 ,w4,c4 ,,preface ,,to ,,! ,,second ,,edition4 ,9 prep>+ ?is edition ,i h endeav\r$ to meet ! requirem5ts ( /ud5ts1 & ( practition]s :o desire to keep up _! hi/ological "w4 ,^? me?ods are select$ :i* h be5 f\nd to "w well 9 practice1 & x has be5 ?\ght bett] to describe a few 9 detail r ?an give a %ort acc\nt ( _m simil> me?ods4 ,i h aga9 to express my obligation to ! v>i\s in/rum5t mak]s = ! illu/rations ( microtomes1 c42 to ,dr4 ,fe>nley1 ( ,brad=d1 = ! description ( 8 me?od = inject+ blood vessels1 & to ,messrs4 ,macmillan & ,co4 = p]mission to copy figures #aj & #aa4 ,w4 ,s4 ,,colman4 ,wimpole ,/reet1 ,w4 ,sept41 #ahif4 ,,cont5ts4 ,,page ,,*apt] ,i4 ,app>atus ,requir$ #a ,,*apt] ,,ii4 ,h>d5+ ,processes #ae ,,*apt] ,,iii4 ,section ,cutt+ #bi ,,*apt] ,,iv4 ,section ,m\nt+ #ee ,,*apt] ,v4 ,g5]al ,/a9+ ,me?ods #fg ,,*apt] ,,vi4 ,special ,me?ods = ,/a9+ ! ,n]ve ,c5tres #hg ,,*apt] ,,vii4 ,special ,me?ods = ,/a9+ ,micro-,organisms & ,blood #ajc ,,*apt] ,,viii4 ,injection ( ,blood ,vessels #abj ,,*apt] ,,ix4 ,directions = ,prep>+ ,individual ,tissues #abi ,index #aec ,,section ,,cutt+ ,,& ,,/a9+4 ,,*apt] ,i4 ,,app>atus ,,requir$4 ,probably !re is no?+ m p]plex+ to a beg9n] ?an to decide :at app>atus is requir$4 ,if he consult a price li/1 x is difficult = him to tell :i* articles w be necess>y1 & :i* w be ei!r luxuries1 or requir$ only = special inve/igation4 ,9 ! foll[+ acc\nt ( requisites1 ^? only w be describ$ :i* x is useful to h always at h&4 ,!y w be f\nd suffici5t = ord9>y "w1 b = special inve/igations a m elaborate equipm5t w be requir$4 ,all /a9+ & o!r reag5ts %\ld be made z f> z possible by ! work] himself1 accord+ to ! directions giv5 9 lat] *apt]s4 ,?is %\ld at any rate be done at fir/1 z ! k ?us ga9$ w prove invaluable4 ,x w also effect a great sav+ if articles t are us$ 9 any quantity1 su* z me?ylat$ _s1 di/ill$ wat]1 c41 are b\ght by ! gallon1 & n 9 small quantities4 ,almo/ all ! processes describ$ "h c be c>ri$ out )\t ! use ( a fully equipp$ laboratory1 9 fact1 9 an ord9>y room4 ,! only furniture requir$ is a firm table1 & a cupbo>d & %elves = /or+ reag5ts4 ,! foll[+ %\ld also be procur$3-- 99,j>s99 or 99bottles991 ) well fitt+ /opp]s or corks1 to conta9 ! tissues :ile be+ h>d5$4 ,!y %\ld n hold less ?an two ounces4 ,empty drug bottles :i* c usually be obta9$ f druggi/s = a few p5ce1 s]ve v well4 ,small] bottles %\ld also be procur$ = keep+ specim5s 9 _s aft] !y h be5 h>d5$ until "o is ready to cut sections4 ,aft] sections h be5 cut f a portion ( ! specim51 ! re/ %\ld be pres]v$1 9 case x is want$ = fur!r inve/igation4 ,ea* specim5 mu/ be labell$1 ) a "n or a numb] correspond+ to a ref]5ce 9 ! note-book1 & a l>ge numb] ( specim5s may !n be kept 9 ! same j>4 ,! be/ way to label !m is to write ! "n or numb] on a piece ( vegetable p>*m5t 9 ord9>y 8m>k+ ink10 & w>m x until ! writ+ is black4 ,! little label %\ld !n be fix$ to a corn] ( ! piece ( tissue ) a /it* or by a f9e p91 & x may be id5tifi$ ye>s aft]w>ds4 ,! importance ( keep+ tissues1 sections1 slides1 c41 99di/9ctly labell$99 _c be too /rongly impress$ on ! beg9n]4 ,! "n1 date1 & o!r p>ticul>s %\ld be inv>iably writt5 on ! label at ! "t4 ,at fir/ ! /ud5t w be incl9$ to neglect ?is1 z he w recognize 8 pieces ( tissue & sections s readily m]ely by _! %ape & g5]al appe>ance4 ,b z "t elapses & simil> specim5s accumulate1 he w f9d x mo/ difficult or ev5 impossible to id5tify "o f ! o!r4 ,a numb] ( #a oz4 & #b oz4 99/opp]$ bottles99 = /a9+ reag5ts4 ,! /opp] ( ^! %\ld be fitt$ ) a rod4 ,?is is done by simply heat+ ! l[] end ( ! /opp] & ! upp] end ( a piece ( glass rod ( suitable l5g? 9 a bl[-pipe1 until !y are pla/ic1 & !n press+ !m toge!r4 99,wat* glasses499--,at lea/ a doz5 wat*-glasses1 9 :i* to p]=m ! op]ations ( /a9+1 cl>ify+1 c4 ,^? ) a flat bottom %\ld be employ$ z !y are less easily upset ?an ! o!rs4 ,pl5ty ( 99filt] pap]s994 ,bo? co>se ones1 = use 9 ! manufacture ( reag5ts1 & small f9e :ite ones 7#b-a_/b in*7 = filt]+ ! /a9+ fluids imm$iately be=e us+ !m1 %\ld be procur$4 ,be=e us+ !m a few drops ( alcohol or di/ill$ wat] %\ld be plac$ 9 !m to saturate ! pap]4 ,?is n only all[s ! fluid to pass ?r\gh m rapidly1 b prev5ts a portion ( x be+ wa/$ ?r\gh be+ absorb$ by ! pores ( ! pap]4 ,sev]al 99ne$les99 m\nt$ 9 h&les4 ,!y mu/ be kept v bright & smoo?1 & c>e mu/ be tak5 t ! po9t does n get turn$ up4 ,a l>ge & small 99funnel994 ,sev]al 99pipettes99 consi/+ ( pieces ( glass tube ) an int]nal diamet] ( #a_/h & ab\t t5 in*es long1 drawn out almo/ to a po9t at "o end4 99,section lift]499--,?is in/rum5t is requir$ = transf]r+ sections f "o reag5t to ano!r1 or f oil ( cloves1 c41 to ! slide4 ,! mo/ conv5i5t =m is ,woodhead's1 made ( ?9 %eet copp]1 :i* all[s ! blade to be b5t at any angle to ! /em4 ,! /em or h&le is ab\t six in*es long1 & cont9u\s )1 & at an angle to x1 a flat blade ab\t #c_/d 94 squ>e ) ! corn]s r\nd$ (f4 ,l>g] ones c be obta9$ = m\nt+ sections ( l>ge size1 e4g41 kidney1 m$ulla oblongata1 c4 ,! surface ( ! blade %\ld be brightly poli%$1 & kept scrupul\sly clean4 ,ord9>y dissect+ 99=ceps994 ,"o or two 99scalpels994 ,a pair ( f9e 99scissors994 ,a 99razor99 or o!r in/rum5t = cutt+ sections4 ,a smoo? 99oil /one99 = keep+ ! razors & knives prop]ly %>p5$4 ,a 99_s lamp99 = w>m+ ! /a9+ fluids4 ,a few 99te/ tubes994 ,a 99m9im measure994 99,scales99 & small weights4 ,a gross ( gr\nd glass slides #c x #a 94 ,half a gross ( gr\nd glass slides #c x #a-a_/b 94 ,half an ounce ( ?9ne/ cov]slips1 #g_/h 94 diamet]4 ,qu>t] ( an ounce ( ?9ne/ cov]slips1 #a-a_/d 94 diamet]4 99,microscope499--,?is is n ! place = a description ( ! microscope z an optical in/rum5t1 b "s h9ts z to ! selection ( "o may be f\nd useful4 ,%[y microscopes ) mu* brass "w %\ld be avoid$1 simplicity ( con/ruction be+ a great recomm5dation4 ,! microscope %\ld h a l>ge heavy base1 ei!r ( ! horse-%oe or tripod patt]n1 l>ge en\gh to af=d a firm base :5 ! microscope is tilt$4 ,me*anical /ages are unnecess>y & !y add greatly to ! exp5se1 & v little to ! utility ( ! in/rum5t = ord9>y hi/ological "w4 ,b9ocul> arrangem5ts also are ( little use = ?is purpose4 ,! microscope %\ld be provid$ ) a 99co>se & f9e adju/m5t991 :i* %\ld be mo/ c>efully te/$ be=e pur*as+ ! in/rum5t4 ,!y %\ld "w freely & smoo?ly1 & ! slighte/ turn 9 ei!r direction %\ld at once alt] ! focus4 ,!re %\ld be a 99rev]sible mirror991 "o side be+ concave & ! o!r plane4 ,! concave surface is ! "o usually employ$1 ! plane surface be+ *iefly us$ 9 conjunction ) ! sub-/age cond5s] = ! exam9ation ( micro-organisms4 ,!re %\ld be an 99eye-piece99 ( mod]ate magnify+ p[]4 ,v p[]ful eye-pieces d n reveal additional details1 b m]ely enl>ge ! image1 & ) x any defects t may be produc$ !re by faults 9 ! objective4 ,eye-pieces ,,ii4 & ,,iv4 ( mo/ mak]s w be ample = mo/ requirem5ts4 99,objectives499--,^! are ! mo/ important p>ts ( ! microscope1 & ! /ud5t w be well advis$ if he sp5ds a little extra money to secure good l5ses4 ,mo/ objectives & /&s are n[ made ) a univ]sal ?read1 s t any objective w fit any make ( /&4 ,_m work]s provide !mselves ) a *eap /& su* z t suppli$ by ,leitz1 & !n fit x ) l5ses by ,zeiss1 or o!r fir/ class mak]4 ,! mo/ useful l5ses are ! #a 94 l[ p[] l5s1 & #a_/e 94 or #a_/f 94 high p[]1 or ,no4 #c & ,no4 #g ( ,cont95tal mak]s1 or ,zeiss's ,a & ,d4 ,a #a_/b 94 l5s w also be f\nd v useful4 ,= m9ute "w1 su* z bact]iology & blood inve/igations1 high] p[]s w be requir$1 #a_/h or #a_/ab 99imm]sion l5ses994 ,^! objectives come extremely close to ! object1 & v ?9 cov] glasses mu/ be employ$4 ,9 ord] to avoid ! refraction caus$ by ! rays trav]s+ ! air betwe5 ! cov]slip & l5s "s imm]sion fluid is plac$ betwe5 ! two4 ,) "s l5ses wat] is employ$1 b usually an oil hav+ ! same refractive index z glass is us$1 & ! "o mo/ g5]ally employ$ is c$> oil 7,zeiss pref]s ! oil f ! species ,junip]us virg9iana74 ,a spot ( oil is plac$ ) a rod j ov] ! object to be exam9$ & ! objective c>efully l[]$ by ! co>se adju/m5t till x comes 9 contact ) ! droplet ( oil4 ,! focuss+ %\ld !n be manag$ ) ! f9e adju/m5t only4 ,:5 ! section has be5 exam9$ ! oil mu/ be remov$ f ! l5s4 ,= ?is purpose a s(t silk h&k]*ief or a special piece ( *amois lea!r may be employ$1 & us$ v g5tly4 ,if all ! oil _c be remov$1 ! h&k]*ief may be moi/5$ ) a little absolute alcohol1 & ! l5s ha/ily wip$4 ,! alcohol mu/ n be all[$ to rema9 9 contact ) ! l5s z x is a solv5t ( ,canada balsam ) :i* ! l5ses are (t5 cem5t$ 9 position4 ,illu/ration3 ,fig4 #a4--;,d\ble or ,triple ,nose-piece4 99,d\ble or triple nose-piece99 7fig4 #a74--,?is me*anical arrangem5t is plac$ on ! l[] end ( ! tube4 ,two or ?ree objectives ( diff]5t magnify+ p[] are atta*$ to x4 ,! nose-piece rotates r\nd a c5tral pivot 9 su* a way t ! objectives c successively be br\ght accurately into position above ! object on ! /age4 ,x is1 !re=e1 a mom5t's "w to replace a high p[] objective by a l[ p[] "o & vice v]sa4 ,x is an extremely conv5i5t "t-sav+ appliance1 & by its use ! risk ( dropp+ & injur+ ! objectives :5 screw+ !m on & (f frequ5tly is avoid$4 ,^? ^: microscopes are n already fitt$ ) ?is appliance c easily h "o fitt$ on at a co/ ( ab\t a sov]eign4 99,sub/age cond5s]499--,?is me*anism = conc5trat+ light on ! object is a necessity = bact]iological "w4 ,! mo/ conv5i5t =m is 99,abbe's illum9at+ app>atus99 7fig4 #b74 ,?is consi/s ( a sy/em ( %ort focus l5ses :i* collects ! light receiv$ by ! mirror1 & ?r[s x on ! object4 ,! am\nt ( light receiv$ f ! mirror is controll$ by an 8iris diaphragm10 ! ap]ture ( :i* c be dilat$ or contract$ by mov+ a small lev] at ! side4 ,x c be fitt$ on to mo/ microscope /&s1 b x is bett] to get a /& 9 ! fir/ in/ance :i* is con/ruct$ to c>ry "o4 ,! co/ ( a microscope v>ies f two gu9eas to two hundr$4 ,!re are _m excell5t microscopes 9 ! m>ket1 & ( ^! sev]al may be m5tion$ :i* ! writ] has f\nd to "w satisfactorily4 ,illu/ration3 ,fig4 #b4--;,abbe's ,illum9at+ ,app>atus4 ,( ! *eap] /ud5t's microscopes ! 8,/>0 microscope made by ,messrs4 ,r4 & ,j4 ,beck1 ( ,cornhill1 ,e4,c41 w be f\nd a safe inve/m5t4 ,x may be obta9$ ) co>se & f9e adju/m5t1 nose-piece1 & #a 94 & #a_/d 94 objectives1 = ab\t #e4 ,^? :o require a bett] in/rum5t w f9d ,beck's 8,pa?ological0 microscope fitt$ ) nose-piece1 ,abbe's illum9ator1 c41 = #af1 meet all requirem5ts4 ,leitz ( ,j5a1 supplies two good & *eap microscopes = #c #ajs4 & #e4 ,!y are n1 h[ev]1 ( uni=m excell5ce1 & !y %\ld be c>efully te/$ by "s compet5t judge be=e ! pur*ase is complet$4 ,leitz imm]sion l5ses are *eap1 & (t5 extremely good1 b %\ld be c>efully te/$ be=eh&1 z _! quality is n q uni=m4 ,! microscopes c be obta9$ f ,mr4 ,a4 ,fraz]1 ,teviot ,place1 ,ed9burgh4 ,! 8,bact]iological0 microscope1 made by ,messrs4 ,swift1 ( ,tott5ham ,c\rt ,road1 is "o ) :i* no "o c be disappo9t$4 ,x is sold ) ,abbe's cond5s]1 triple nose-piece1 #a_/f 941 & a #a_/ab 94 imm]sion objective1 = j "u #bj4 ,bo? /& & l5ses are turn$ out 9 ,swift's fir/-class /yle1 & ^? :o c af=d ! initial outlay w n regret x4 ,or ! /& may be pur*as$1 & ! objectives & accessories add$ s+ly f "t to "t4 ,among ,cont95tal mak]s1 excell5t microscopes = hi/ological "w are turn$ out by _m mak]s4 ,zeiss's l5ses /& des]v$ly high 9 reputation1 z no faulty ,zeiss l5s ev] leaves ! works1 & _! optical prop]ties are ne>ly p]fect4 ,= ?is gu>antee1 h[ev]1 ! pur*as] has to pay some:at high] prices1 b ! money is well inve/$4 ,zeiss's ag5cy is at #bi ,m>g>et ,/reet1 ,reg5t ,/reet1 ,w4 ,rei*]t1 ( ,vi5na1 sells microscopes & l5ses :i* are modell$ on ! l9es ( ^? ( ,zeiss1 & ?\gh *eap] are (t5 equal to !m 9 excell5ce1 b ! quality is n q uni=m4 ,8 in/rum5ts c be obta9$ ?r\gh any optician1 b 8 ag5t 9 ?is c\ntry is ,mr4 ,a4 ,fraz]1 ,teviot ,place1 ,ed9burgh4 ,be=e buy+ a microscope ! /ud5t %\ld obta9 an illu/rat$ price li/ f any ( ! firms m5tion$ above1 &1 hav+ select$ an in/rum5t1 he %\ld te/ x v c>efully1 or bett] get "s exp]i5c$ fri5d to te/ x = him1 be=e decid+ to pur*ase x4 ,delicate te/ objects su* z diatoms1 scales ( butt]fly's w+1 or a /a9$ specim5 ( micro-organisms %\ld be employ$4 ,! co>se & f9e adju/m5ts %\ld be tri$4 ,!y %\ld "w freely & smoo?ly & )\t any delay4 ,! def9ition ( ! l5s mu/ be te/$ ) ! f9e objects m5tion$4 ,! field %\ld be q flat1 i4e41 e "p %\ld be 9 focus at ! same "t1 & ! def9ition %\ld be p]fectly %>p & accurate1 & ! te/ objects )\t d\ble cont\r4 ,! field %\ld be totally free f prismatic col\rs4 ,if !re is a halo ( col\r ar\nd ! objects x indicates a defect 9 ! optical prop]ties ( ! objective1 & ano!r %\ld be select$4 ,a microscope mu/ always be treat$ ) ! greate/ c>e4 ,j>s & falls t5d to slightly loos5 & %ift ! l5ses1 & to p]man5tly impair its optical prop]ties4 ,du/ mu/ be mo/ c>efully exclud$4 ,?is is be/ effect$ by keep+ ! in/rum5t "u a glass bell j> :5 n 9 use4 ,! l5ses %\ld be wip$ z little z possible1 & :5 x is necess>y1 v s(t *amois lea!r %\ld be employ$4 ,! microscope mu/ be kept 9 a dry room1 or ! brass "w w soon t>ni% & ! /eel p>ts w t5d to ru/4 ,,*apt] ,,ii4 ,,h>d5+ ,,processes4 ,= ! satisfactory exam9ation ( tissues x is necess>y t !y %\ld be 8h>d5$0 9 c]ta9 fluids4 ,! object ( ?is is to give ! specim5s great] consi/5ce1 s t ?9 sections may be m readily obta9$ & m safely manipulat$1 & also to 8fix0 ! tissue elem5t z f> z possible 9 ! same relative position z 9 ! liv+ body4 ,! h>d5+ process also acts on ! protoplasm ( ! cells1 & prev5ts _! swell+ up :5 plac$ 9 wat]1 & 9 ! v>i\s /a9+ fluids4 ,! fluid us$ mu/ be "o :i* w n itself injure ! specim51 & :i* c be ?or\ghly remov$ by wa%+1 s t x may n int]f]e ) /a9+ op]ations4 ,! specim5s %\ld be kept :ile h>d5+ 9 wide m\!d bottles1 on ! bottom ( :i* a little cotton wool or t[ has be5 laid4 ,?is all[s ! h>d5+ fluid to come freely 9 contact ) ! "u surface ( ! pieces ( tissue1 & prev5ts _! be+ flatt5$ aga9/ ! h>d glass bottom4 ,! h>d5+ fluid requires *ang+ occasionally4 ,?is %\ld always be done at ! end ( tw5ty-f\r h\rs1 9 ord] to get rid ( any deposit ( blood1 c41 t may h accumulat$4 ,besides ?is1 ! tissue :5 plac$ 9 ! fluid conta9$ a good deal ( wat] :i* w h dilut$ x & consequ5tly an e>ly *ange is desirable4 ,aft]w>ds ! fluid requires to be *ang$ only z (t5 z x becomes turbid1 or any deposit occurs1 usually ab\t once a week4 ,:ile h>d5+1 specim5s %\ld be kept 9 a cool place1 z w>m? fav\rs *anges 9 ! cells1 c4 ,9 manipulat+ ! portions ( organs1 =ceps %\ld always be us$ & ^! ) great g5tl5ess4 ,! specim5s %\ld nev] be impal$ ) ne$les1 or unsightly holes1 :i* may ev5 be mi/ak5 = pa?ological appe>ances1 w appe> :5 a section is exam9$ "u ! microscope4 ,x requires "s practice to "k :5 ! tissue is suffici5tly h>d5$4 ,! object aim$ at is to make !m n really h>d b t\gh4 ,x is almo/ unnecess>y to add t 9 te/+ ?is ) ! f+]s ! utmo/ g5tl5ess mu/ be obs]v$1 or s]i\s damage may be done to ! tissue4 ,:5 ! tissue is suffici5tly h>d5$ ! h>d5+ fluid mu/ be ?or\ghly dissolv$ out4 ,?is is mo/ quickly effect$ by plac+ ! specim5 9 a bas9 into :i* cold wat] f a tap is con/antly runn+4 ,! tissue may !n be remov$ 7=ceps always be+ us$ & nev] ! ne$le7 & plac$ 9 an imb$d+ m$ium z subsequ5tly direct$2 or1 if x is n to be cut at once1 into equal p>ts ( me?ylat$ _s & wat]1 9 :i* x may be kept indef9itely1 ! fluid be+ *ang$ if x becomes at all cl\dy4 ,x is unnecess>y = ord9>y "w to h m ?an ! foll[+ h>d5+ fluids3--99,mll]'s fluid993-- ,potassium ,bi*romate #b-a_/d grms4 #c-a_/b dra*ms4 ,sodium ,sulphate #a grm4 #a-a_/b dra*ms4 ,wat] to #ajj c4c4 #a p9t4 ,two dra*ms ( c>bolic acid are sometimes add$ to ea* p9t ( ! fluid b z a rule x is n necess>y4 ,mll]'s fluid is ! mo/ g5]ally useful ( ! v>i\s fluids employ$1 = ! foll[+ reasons3-- #a4 ,x causes v little %r9k+ ( ! elem5ts ( ! tissue1 & h5ce may be employ$ = mo/ delicate objects1 e4g41 ! ret9a & embryos4 #b4 ,9 consequ5ce ( its n mak+ ! tissues %r9k1 x does n squeeze ! blood out ( ! vessels & :]e ! organ has be5 conge/$ be=e dea?1 we may1 by us+ ,mll]'s fluid1 pres]ve a natural injection ( ! capill>ies4 #c4 ,!re is comp>atively little dang] ( ov]-h>d5+ ! tissue & r5d]+ x brittle4 #d4 ,sections ( organs h>d5$ 9 ,mll]'s fluid are usually firm & easy to manipulate4 ,!y d n t5d to curl up or adh]e to "o ano!r z mu* z ^? h>d5$ 9 _s4 #e4 ,x readily p]meates ! tissues1 & h5ce l>ge portions ( organs1 or ev5 ! entire organ may be satisfactorily h>d5$ 9 x4 #f4 ,x is v *eap4 ,a gallon c be made up = ab\t eightp5ce4 ,! fluid has h[ev] c]ta9 slight drawbacks3-- #a4 ,! h>d5+ process is a sl[ "o occupy+ f\r to eight weeks4 #b4 ,! fluid gives a p]man5t d+y col\r to ! tissue4 ,?is does n cause any inconv5i5ce = microscopic purposes1 b x is a disadvantage :5 x is int5d$ to pres]ve ! re/ ( ! specim51 z a nak$ eye prep>ation4 ,9 su* cases ! organ %\ld be h>d5$ 9 _s1 c>bolic acid1 or =mal4 ,mll]'s fluid c be us$ = almo/ any tissue4 ,x is especially useful = ^? :i* conta9 a l>ge quantity ( fluid1 or ( blood1 & is ess5tial = n]ve tissues :i* x is int5d$ to /a9 by ,pal's me?od 7p4 #hi74 ,to h>d5 a specim5 9 x at lea/ tw5ty times ! bulk ( fluid mu/ be employ$4 ,! fluid mu/ be *ang$ on ! ?ird day1 & aft]w>ds ab\t e week z may be requir$4 99,me?ylat$ _s99 is a v useful h>d5+ ag5t4 ,x h>d5s 9 "o to ?ree weeks accord+ to ! size ( ! tissue & ! quantity ( _s us$4 ,its disadvantages are3-- #a4 ,x is m apt to ov]h>d5 ?an ,mll]'s fluid4 #b4 ,x causes a great deal ( %r9k+ ( ! tissue & ?us squeezes mu* ( ! blood out ( ! vessels4 ,x is mo/ useful 9 h>d5+ tissues conta9+ mu* epi!lium1 e4g41 kidney1 epi!lioma1 c4 ,_s is also frequ5tly employ$ to complete ! h>d5+ by ,mll]'s fluid & to pres]ve tissues aft] !y h be5 h>d5$4 ,ab\t t5 or fifte5 times ! bulk ( _s %\ld be us$ = "o ( ! tissues4 ,! fluid %\ld be *ang$ on ! ?ird day & aft]w>ds z requir$4 99,mll]'s fluid & _s499--,?is is a useful comb9ation = _m purposes4 ,x is made ?us3--,mll]'s fluid1 ?ree p>ts2 me?ylat$ _s1 "o "p4 ,! fluid mu/ be all[$ to cool aft] mix+ be=e be+ us$1 & if necess>y filt]$4 ,x w h>d5 specim5s satisfactorily 9 ?ree weeks4 99,mll]'s fluid & =mal499--,is an extremely useful mixture made by add+ "o "p ( =mal to n9e ( ,mll]'s fluid4 ,x h>d5s 9 a mu* %ort] "t ?an ,mll]'s fluid & causes v little %r9kage4 99,absolute alcohol499--,us$ z a h>d5+ ag5t :]e ! tissues are to be exam9$ = micro-organisms1 & = specim5s to be /a9$ by ,nissl's me?od 7p4 #aja74 ,a *eap] & equally effective h>d5+ m$ium is made by dehydrat+ me?ylat$ _s by add+ "o ounce ( fus$ c>bonate ( potassium to ea* p9t ( me?ylat$ _s1 & decant+4 ,small pieces mu/ be us$4 ,! dep?s ( ! block %\ld n exce$ #c_/h in*4 ,! fluid %\ld be *ang$ on ! ?ird day4 ,h>d5+ w be complet$ 9 ab\t t5 days or ev5 e>li]4 99,osmic acid499--,= rapidity ( action1 & = rapid fix+ ( all ! tissue elem5ts 9 _! natural position osmic acid is "o ( ! be/ h>d5+ reag5ts we possess4 ,its disadvantages z a h>d5+ ag5t are3-- #a4 ,its exp5se4 #b4 ,its irritat+ & corrosive vap\r4 #c4 ,! fact t only small pieces ( tissue c be h>d5$ 9 x1 s9ce ! ext]nal surface is v rapidly h>d5$ & ?us ! fluid is prev5t$ f p5etrat+ into ! c5tre ( ! lump4 ,x is mo/ frequ5tly us$ z a h>d5+ ag5t = v delicate /ructures1 su* z ! ret9a1 or embryos1 or = fre% sections ( bra9 7p4 #ie74 ,! acid itself may be procur$ 9 seal$ tubes1 ea* conta9+ "o gramme4 ,^! %\ld be brok5 9 a bottle "u suffici5t di/ill$ wat] to make a "o p] c5t4 solution4 ,! bottle conta9+ x %\ld be cov]$ ) br[n pap] to exclude ! light4 ,= h>d5+ purposes small pieces ( ! tissue1 n mu* l>g] ?an a pea1 %\ld be plac$ 9 ! acid1 ! "o p] c5t4 solution be+ dilut$ ) five to t5 volumes ( di/ill$ wat]4 ,! tissues may be left 9 ?is = f ?ree to five days4 ,!y mu/ !n be 99?or\ghly99 wa%$ 9 di/ill$ wat] & may aft]w>ds be pres]v$ 9 me?ylat$ _s4 ,bo? ! h>d5+ & ! subsequ5t wa%+ mu/ be c>ri$ on 999 ! d>k994 ,osmic acid is also a mo/ valuable /a9+ reag5t 7see p4 #ha74 99,c>bolic acid99 7#e p] c5t474--,may be us$ to h>d5 almo/ any tissue1 b is p>ticul>ly useful = h>d5+ n]v\s tissues su* z bra9 or sp9al cord :i* are aft]w>ds to be pres]v$ z museum specim5s4 ,x does n dis*>ge ! col\r ( a specim5 s rapidly z _s4 ,?ree or f\r times ! bulk ( fluid %\ld be us$4 ,x requires *ang+ at ! end ( tw5ty-f\r h\rs1 & aga9 at ! end ( ! fir/ week4 ,saturat$ aque\s solution ( 99corrosive sublimate99 is "o ( ! mo/ conv5i5t h>d5+ reag5ts = small pieces ( delicate tissue1 e4g41 embryos4 ,x h>d5s !m 9 a few days4 ,:5 !y are suffici5tly h>d5$ ! m]curial salt %\ld be remov$ by wa%+ fir/ 9 me?ylat$ _s = a few h\rs & !n 9 runn+ wat]4 99,=mal499--,an aque\s solution conta9+ ab\t #ce p] c5t4 ( =maldehyde4 ,x is a rapid h>d5+ ag5t1 causes v little %r9kage ( ! tissues1 & does n dis*>ge ! col\r ( ! specim5s z mu* z alcohol4 ,= h>d5+ =mal %\ld be us$ z a two to five p] c5t4 solution 9 di/ill$ wat]4 ,x may also be us$ z a t5 p] c5t4 solution = m\nt+ museum prep>ations1 b !re is "s t5d5cy = a cl\dy deposit to =m on ! glass aft] a "t4 ,x is ! mo/ suitable h>d5+ ag5t at our disposal = eyes4 ,x rapidly fixes ! tissue elem5ts1 b does n cause mu* contraction4 ,x may also be us$ = h>d5+ ! bra9 & sp9al cord4 ,l>ge quantities ( fluid mu/ be us$ = ! latt] purpose & x mu/ be frequ5tly *ang$4 ,z soon z !y are suffici5tly h>d5$ !y %\ld be transf]r$ to me?ylat$ _s4 99,m>*i's fluid499--,?is consi/s (3-- ,mll]'s fluid #b p>ts4 ,osmic acid solution 7"o p] c5t47 #a "p4 ,x is us$ = h>d5+ specim5s z a prelim9>y to ,golgi's me?od = /a9+ n]ve cells 7p4 #ig71 & also to complete ! h>d5+ ( sections ( sp9al cord1 c41 be=e employ+ ,s*f]'s modification ( ! ,weig]t ,pal hmatoxyl9e me?od 7p4 #ia74 ,x is also us$ z a /a9 = rec5tly deg5]at$ n]ve tracts & fibres1 especially aft] exp]im5tal lesions4 ,! fluid has little p5etrat+ p[]1 & !re=e tissues mu/ be cut into small pieces1 ab\t #c_/h in* cube4 ,x is n necess>y to place !m 9 ?is fluid at once on removal f ! body1 b ! prelim9>y h>d5+ mu/ be 9 ,mll]'s fluid & n 9 alcohol1 c4 ,,special ,,h>d5+ ,,reag5ts ,,= ,,rapid ,,fixation ,,9 ,,ord] ,,to ,,/udy ,,cell ,,/ructure4 #a4 99,alcohol499 #b4 99,flemm+'s solution99 7modifi$ by ,fri$mann73-- ,osmic acid 7"o p] c5t47 #c c4c4 7xxx474 ,glacial acetic acid #b c4c4 7xx474 ,*romic acid 7"o p] c5t47 #db c4c4 7j47 ,small pieces %\ld be h>d5$ 9 ?is fluid = twelve to tw5ty-f\r h\rs1 & !n wa%$ & transf]r$ to alcohol = "s days be=e /a9+4 #c4 99,nitric acid499--,a t5 p] c5t4 solution 9 di/ill$ wat]4 ,x h>d5s ! tissue 9 ?ree to f\r h\rs1 & %\ld be foll[$ by #gj p] c5t4 alcohol1 ! h>d5+ be+ complet$ 9 absolute alcohol4 ,9 us+ any ( ^! me?ods x is necess>y t ! tissue be remov$ f ! body dur+ life or imm$iately aft] dea?4 ,!y are employ$ = reveal+ ! *anges 9 ! cells & _! nuclei 9 rapidly gr[+ or inflam$ tissue1 = /udy+ k>yok9esis 9 canc] cells1 & inve/igat+ ! appe>ance ( n]ve cells & gl& cells at re/1 :5 actively employ$ & :5 fatigu$2 & !y are also mo/ useful 9 prep>+ specim5s ( ! 8p>asitic bodies0 :i* h be5 describ$ 9 _m canc] cells4 ,,decalcify+ ,,fluids4 ,us$ 9 ! prep>ation ( bone1 too?1 osse\s tum\rs1 c4 ,! two be/ fluids = g5]al use are3-- 99,*romic & nitric fluid499--,?is is made z foll[s3-- ,*romic acid #a gramme #de gra9s4 ,nitric acid #b grammes #a-a_/b dra*ms4 ,wat] #bjj c4c4 #a p9t4 ,if ! bone is n v compact ! fluid may be us$ dilut$ ) an equal quantity ( wat]4 ,a l>ge quantity ( fluid %\ld be us$1 & l all decalcify+ fluids1 x %\ld be frequ5tly *ang$4 ,z soon z ! specim5 is suffici5tly flexible1 x %\ld be ?or\ghly wa%$ 9 runn+ wat] = "s h\rs1 & !n transf]r$ to _s until x is conv5i5t to cut sections4 99,von ,ebn]'s solution993-- ,hydro*loric acid #a gramme #a-a_/b dra*ms4 ,common salt #aj grammes #b ounces4 ,wat] to #ajj c4c4 #a p9t4 ,x is a v useful decalcify+ ag5t1 b causes ! fibr\s elem5ts to swell up r m ?an *romic & nitric fluid4 ,a l>ge quantity ( ! fluid mu/ be us$1 & x %\ld be *ang$ daily4 ,x mu/ be v ?or\ghly wa%$ out 9 runn+ wat] :5 ! decalcification is complet$4 99,blea*+ solution99 7eau de ,javelle74 7#a7 8,*loride ( lime0 7blea*+ p[d]7 #bj #a_/b oz4 ,wat] #ajj #b-a_/b oz4 ,%ake up well4 7#b7 ,c>bonate ( potassium #bj #a_/b oz4 ,wat] #ajj #b-a_/b oz4 ,mix ! two solutions4 ,all[ !m to /& = an h\r & filt]4 ,x is us$ *iefly = cle>+ vegetable sections b may also be us$ = sections conta9+ a l>ge quantity ( pigm5t4 ,x is p>ticul>ly useful 9 decol\riz+ sections ( 8madura foot0 due to ! pres5ce ( a black fungus4 ,,*apt] ,,iii4 ,,section ,,cutt+4 99,emb$d+ ( sections499--,be=e sections are made ! tissues require to be emb$d$ 9 "s fluid1 :i* w p]meate _! int]/ices1 & is capable ( be+ r5d]$ firm s z to support ! mo/ delicate p>ts :5 ! knife passes ?r\gh ! tissue4 ,! mo/ g5]ally useful sub/ances are3-- 7#a7 gum1 7#b7 celloid91 7#c7 p>aff9 or wax4 99,gum499--,pick$ col\rless gum arabic #b p>ts1 cold wat] #c p>ts4 ,leave ) frequ5t /irr+ until dissolv$4 ,add t5 drops ( c>bolic acid to ea* ounce ( ! mucilage4 ,specim5s are ?or\ghly fre$ f all trace ( ! h>d5+ fluid by wa%+ 9 wat]1 & ! tissue is !n plac$ 9 ! gum solution = at lea/ twelve h\rs1 or if en\gh c>bolic acid be add$1 x may be left !re = an indef9ite "t4 ,:5 froz51 gum =ms a firm non-cry/all9e mass1 :i* supports ! tissue on all sides4 ,x mu/ n be froz5 too deeply1 or x becomes h>d & r brittle & is apt to injure ! razor4 ,if ?is h occurr$ ! surface c be s(t5$ suffici5tly by brea?+ g5tly on x4 ,aft] cutt+ 9 gum1 ! sections are g5tly remov$ f ! knife into di/ill$ wat] by a s(t camel's hair bru%1 & left !re = an h\r or two1 until ! m$ium is entirely dissolv$ out4 ,!y may !n be /a9$ & m\nt$1 or !y may be put away 9 _s = an indef9ite "t1 & !n /a9$ & m\nt$4 99,celloid999 is = _m purposes almo/ an ideal emb$d+ m$ium4 7#a7 ,x has great p5etrat+ p[]2 7#b7 x c be made ( an admirable consi/5ce = cutt+ purposes2 7#c7 aft] sections are made x all[s !m to be v freely manipulat$ )\t fe> ( injur+ !m3 7#d7 & be+ p]fectly transp>5t & homog5e\s 9 ?9 sections1 x does n require to be remov$ f a section be=e m\nt+4 ,x is insoluble 9 wat]1 & 9 weak _s2 slightly soluble 9 alcohol ( m ?an #ij p] c5t4 /r5g?1 & v readily soluble 9 e!r1 or 9 a mixture ( alcohol & e!r4 ,! la/ solv5t is ! "o commonly employ$4 ,! emb$d+ solution is made ?us3-- ,take "s pure celloid9 78,s*]+'s10 sold 9 boxes conta9+ an ounce ( %av+s1 is v good7 & p\r on x ab\t eight times its volume ( a mixture ( equal p>ts ( absolute alcohol & e!r4 ,all[ ?is to /& all night until ! celloid9 is dissolv$4 ,! solution %\ld be made ab\t ! consi/5ce ( ord9>y mucilage4 ,x is also conv5i5t to h a ?9n] solution made by us+ d\ble ! proportion ( alcohol & e!r4 ,bo? solutions %\ld be kept 9 wide m\!d /opp]$ bottles1 & ! /opp] %\ld be well greas$ ) vasel9e z an additional ob/acle to ! evaporation ( ! volatile e!r4 ,be=e emb$d+ a specim5 x is necess>y to dehydrate x ?or\ghly = twelve to tw5ty-f\r h\rs 9 absolute alcohol4 ,x %\ld !n be plac$ 9 a mixture conta9+ alcohol & e!r = an h\r or two1 & aft]w>ds transf]r$ to ! ?9 solution ( celloid9 = tw5ty-f\r h\rs1 & !n to ! ?ick solution = ! same p]iod4 ,! celloid9 p5etrates sl[ly & 9 ! case ( n]ve tissues & o!r delicate /ructures x is wise to give ! full all[ance ( "t = ! diff]5t /eps4 ,:5 ! tissue has be5 ?or\ghly p]meat$ by ! celloid91 x is g5tly remov$ f ! celloid9 & plac$ 9 position on a piece ( cork ( suitable size = clamp+ 9 ! hold] ( ! microtome4 ,celloid9 is pa9t$ r\nd ! object s t x is support$ on e side4 ,x is !n left expos$ to ! air until ! surface has become firm1 :5 ! cork is plac$1 ) ! tissue d[nw>ds1 9 me?ylat$ _s4 ,! cork floats b ! tissue & celloid9 rema9 subm]g$4 ,at ! end ( tw5ty-f\r h\rs ! celloid9 w h become semi-opaque & opalesc5t1 & ( ! same consi/5ce z h>d boil$ :ite ( egg4 ,:5 x is impossible to wait s long1 rapid h>d5+ ( ! celloid9 may be secur$ by imm]s+ x 9 me?ylat$ *lor(orm 9 place ( _s1 b ! sl[] me?od gives m uni=mly satisfactory results4 ,pieces ( tissue emb$d$ 9 celloid9 may also be cut on a freez+ microtome4 ,aft] ! celloid9 has become firm by imm]sion 9 me?ylat$ _s1 ! tissue ) ! celloid9 r\nd x may be cut (f ! cork1 wa%$ 9 wat] to remove ! alcohol1 & !n soak$ = an h\r or two 9 gum1 plac$ on ! plate ( an e!r spray microtome1 froz5 & cut 9 ! usual way4 ,subsequ5t /a9+ op]ations are conduct$ 9 ! same way z = sections cut by h& or 9 gum4 ,z celloid9 is only slightly /a9$ by hmatoxyl91 alum c>m9e1 borax c>m9e1 c41 x is n necess>y to remove x f ! sections1 b x exhibits s int5se a /a9+ reaction ) anil9e dyes t x is necess>y to remove x by treatm5t ) alcohol & e!r ei!r be=e or aft] ! /a9+ op]ation4 ,! sections aft] /a9+ may be m\nt$ 9 ,f>rant's solution 7p4 #ei71 or 9 ,canada balsam 7p4 #fa74 ,if ! latt] m$ium is employ$1 ! section %\ld be cl>ifi$1 aft] dehydration 9 alcohol1 by means ( oil ( b]gamot1 or oil ( origanum1 in/ead ( oil ( cloves1 z ! latt] dissolves out ! celloid9 & causes ! section to break up4 ,celloid9 is mo/ useful = cutt+ sections ( ! coats ( ! eye1 ( ! int]nal e>1 & ( bone m>r[4 ,x %\ld always be us$ = ! ,weig]t-,pal hmatoxyl9e me?od ( /a9+ ! n]v\s c5tres1 z x protects ! section f be+ injur$ by ! transf]5ce f "o fluid to ano!r :i* is repeat$ly requir$ dur+ ! process4 ,! /a9 is completely dis*>g$ f ! celloid9 by ! decol\ris+ solution us$ 7p4 #ij74 ,p>aff94--,p>aff9 is a v conv5i5t emb$d+ m$ium = delicate /ructures1 z v ?9 sections c be obta9$ & ! p>aff9 ne$ n be remov$ f ! section until ! latt] is safely on ! slide4 ,x is unsuitable = l>ge sections4 ,/a9+ op]ations are n easily c>ri$ out aft] cutt+ 9 p>aff91 & x is bett] to /a9 ! blocks ( tissue 9 bulk be=e emb$d+4 ,! be/ /a9s = p5etrat+ are borax c>m9e 7p4 #ge71 alum c>m9e 7p4 #gf71 & ,kle95b]g's hmatoxyl9e 7p4 #gj74 ,! tissue mu/ be left 9 !m = f\r to t5 days4 ,v>i\s k9ds ( p>aff9 are employ$4 ,x is usual to keep two k9ds1 "o 8s(t10 melt+ at #aaj ,f41 & ano!r 8h>d10 melt+ at #adj ,f4 ,a mixture ( two p>ts ( ! h>d & "o ( ! s(t w be f\nd mo/ g5]ally useful4 ,9 w9t] a l>ge proportion ( ! s(t v>iety & 9 hot wea!r a l>g] proportion ( ! h>d may be requir$4 ,a p>aff9 mass :i* is always available has be5 sugge/$ rec5tly by ,dr4 ,f4 ,e4 ,batt51 :o employs an ord9>y :ite c&le1 compos$ ( p>aff9 & wax4 ,if ! mass is f\nd to be too h>d1 x c easily be made ( a suitable consi/5ce by add+ a little p>aff9 ) a l[ melt+ po9t4 ,to prep>e a piece ( tissue = emb$d+ 9 p>aff91 x %\ld be /a9$1 wa%$ 9 di/ill$ wat]1 & z mu* moi/ure z possible remov$ by blott+ pap]4 ,! block is !n dehydrat$1 fir/ 9 me?ylat$ _s = sev]al h\rs1 f9ally 9 absolute alcohol4 ,x is tak5 c>efully by means ( =ceps f ! alcohol & plac$ 9 xylol = an h\r or two accord+ to size4 ,sup]flu\s xylol is remov$ f ! surface1 & ! tissue plac$ 9 ! melt$ p>aff94 ,?is w set r\nd ! cold tissue at once1 b soon melts aga9 & mu/ be kept at a temp]ature j above melt+ po9t = "o to f\r h\rs1 accord+ to size4 ,! tissue is !n transf]r$ to a m\ld 7:i* c be easily made ( pap]71 ab\t half an in* cube1 & melt$ p>aff9 p\r$ r\nd x until ! m\ld is full4 ,! m\ld may be made by fold+ a piece ( pap] to =m a box ab\t half an in* cube1 or a small pill box may be us$4 ,ano!r conv5i5t me?od is to place two ,l-%ap$ pieces ( lead 9 contact ) ea* o!r s z to enclose a space ( suitable size z 9 ! diagram 7fig4 #c74 ,! tissue is n[ h]metically seal$1 & c be kept indef9itely if x is n conv5i5t to cut x at ! "t4 ,to prep>e x = cutt+1 all sup]flu\s p>aff9 is trimm$ away ) a w>m knife1 & ! block is fix$ on a piece ( wood1 cut s z to suit ! clamp ( ! microtome1 by melt+ ! l[] end ( ! p>aff9 block ) a hot ne$le or wire & press+ x d[n on ! wood4 ,illu/ration3 ,fig4 #c4 ,:5 sections are cut !y may be transf]r$ s+ly to ! slide 7:i* %\ld be lightly sme>$ be=eh& ) a saturat$ solution ( celloid9 9 oil ( cloves71 or !y may be cut s t ! back ( "o section ( ! p>aff9 block adh]es to ! front ( ! next1 & 9 ?is way a cont9u\s delicate ribbon ( s]ial sections is obta9$4 ,! ribbon is brok5 up into l5g?s ( ab\t two & a half in*es & transf]r$ to ! slide1 on :i* sev]al ribbons may be plac$ side by side1 & s a l>ge numb] ( sections kept 9 ! ord] 9 :i* !y are cut4 ,a m>k %\ld be made on ! slide to indicate :]e ! s]ies beg9s1 & ea* slide %\ld be numb]$1 s t ! exact position ( ea* section 9 ! s]ies c be recognis$ at once4 ,be=e m\nt+1 ! p>aff9 mu/ be remov$ f ! sections4 ,?is is easily done on ! slide 9 ! case ( s+le sections & ( ribbons4 ,if ! sections are curl$1 a little w>m? w make !m unb5d & lie flat4 ,! slide is w>m$ ov] a _s lamp until ! p>aff9 j melts4 ,! sections w keep _! places ow+ to ! celloid9 b5ea?4 ,xylol is !n all[$ to fl[ ov] ! slide f a pipette1 until ! p>aff9 has be5 completely dissolv$1 :i* c be asc]ta9$ by glanc+ at ! sections "u ! l[ p[] ( ! microscope4 ,! slide is plac$ 9 an almo/ v]tical position to let ! xylol dra9 (f1 excess is wip$ (f f ! edge ( ! slide ) blott+ pap]1 a drop ( ,canada balsam solution 7p4 #fa7 is run on ! slide1 & a cov]-glass ( suitable size is appli$4 99,microtomes499--,aft] a l>ge am\nt ( practice1 p]sons ) a fair am\nt ( manual dext]ity may acquire suffici5t skill to be able to cut v satisfactory sections ( specim5s emb$d$ 9 p>aff91 c41 by h&4 ,9 ! ,pa?ological ,laboratory ( a l>ge ,g]man ,univ]sity1 until q rec5tly ! use ( a microtome 0 prohibit$ by ! ,pr(essor1 :o is himself a mo/ di/+ui%$ hi/ologi/4 ,! am\nt ( "t exp5d$ be=e "o acquires ! necess>y skill1 & ! *eapness & great conv5i5ce ( ! mod]n microtome h comb9$ to ?r[ h& cutt+ into ! backgr\nd1 & "s =m ( microtome is n[ almo/ univ]sally adopt$4 ,illu/ration3 ,fig4 #d4--;,ca?c>t's ,e!r ,spray ,microtome4 ,a1 ,b4 ,wood5 frame & supports4 ,c4 ,glass runn]s4 ,g4 ,screw = rais+ ! z9c plate ,h4 ,j4 ,e!r bottle4 ,l4 ,tube f air bell[s4 ,( ^! !re are a v l>ge numb] 9 ! m>ket1 ea* hav+ special advantages1 & (t5 special drawbacks4 ,a few ( ! m g5]ally useful only w be describ$4 ,we h microtomes = cutt+ 9 gum froz5 by e!r spray or ice1 & ^? int5d$ = cutt+ 9 p>aff9 or celloid94 ,ca?c>t's 99e!r spray microtome99 7fig4 #d74--,?is1 or its m rec5t modifications 7see lat]71 is p]haps ! mo/ useful & economical microtome = ! purposes ( ! /ud5t4 ,its prime co/ is l[1 x is small & portable z well z be+ clean & inexp5sive to "w )4 ,x consi/s ( an oak frame :i* c be firmly clamp$ on to a table4 ,on ?is frame are two n>r[ p>allel supports ab\t two in*es high1 :i* are cov]$ by /rips ( plate glass1 & s]ve z smoo? re/s along :i* ! razor may glide 9 mak+ sections4 ,betwe5 !m is a brass well & 9 ?is a z9c plate firmly fix$ 9 ! horizontal position1 :i* is almo/ at ! level ( ! glass runn]s4 ,x is capable ( be+ rais$ or l[]$ ?r\gh ab\t #c_/h in* by means ( a screw ) a v f9e & accurate ?read4 ,?is screw is turn$ by a l>ge mill$ :eel b5ea? ! microtome4 ,j b5ea? ! z9c plate are two small tubes1 "o connect$ ) an india-rubb] bell[s1 ! o!r ) a bottle at ! side :i* conta9s e!r4 ,z ! air issues f ! fir/ tube1 x passes ov] ! op5 end ( ! second1 & ?us draws ! e!r out & makes x play on ! z9c plate1 & at ! same "t causes x rapidly to evaporate1 & s r$uces ! temp]ature ( ! z9c plate4 ,9 cutt+ sections ) ?is microtome ! tissue is tak5 out ( ! gum & plac$ on ! z9c plate4 ,! bell[s are !n work$ until ! gum on ! z9c plate is completely froz54 ,! plate %\ld be l[]$ by means ( ! screw until ! surface ( ! piece ( tissue is on a level ) ! glass runn]s4 ,^! & ! razor %\ld !n be wett$ ) wat]4 ,! razor be+ held firmly 9 ! h& is pu%$ along ! glass runn]s 9 a r oblique direction4 ,! plate %\ld !n be rais$ by turn+ ! screw bel[ ?r\gh a v small arc1 ano!r section tak5 (f & s on4 ,sections are c>efully remov$ f ! razor to a vessel ( wat] by means ( a s(t wet camel's hair bru%4 ,! ne$le %\ld nev] be us$ = ?is purpose4 ,if ! specim5 is v delicate1 & likely to be spoil$ by be+ curl$ up on ! knife1 ! latt] %\ld be kept cold by frequ5tly dipp+ x 9 a vessel conta9+ lumps ( ice 9 wat]4 ,! gum w !n rema9 froz5 aft] cutt+1 & support ! tissue bett]4 ,ea* section %\ld be at once transf]r$ to a glass slide f ! knife1 wa%+ x (f ) a /ream ( ice wat] f a pipette4 ,! knife t is us$ may be an ord9>y razor1 ) ! edge gr\nd /raight4 ,x requires to be held /eadily ) bo? h&s4 ,z ?is is r inconv5i5t1 ,dr4 ,%]idan ,dlp9e sugge/$ ! employm5t ( an ord9>y plane iron su* z is us$ 9 a c>p5t]'s plane4 ,?is only requires "o h&1 & ! o!r c be kept on ! head ( ! screw b5ea? to raise ! plate at once aft] ea* /roke ( ! knife4 ,its disadvantages are t x is r heavy = prolong$ work+1 & t x is less easy to 8set0 ?an a razor4 ,a4 ,fraz] has rec5tly introduc$ a valuable improvem5t 9 ! ,ca?c>t microtome 7fig4 #e74 ,9 ?is ! brass frame c>ry+ ! z9c plate & e!r spray tubes is surr\nd$ by a brass cyl9d]1 9 :i* x fits accurately1 & is pu%$ up z desir$ by turn+ ! screw b5ea? ! in/rum5t4 ,?is brass frame & ) x ! z9c plate1 c41 c be easily drawn altoge!r out ( ! out] tube1 & replac$ by a second brass well1 :i* exactly fits its place & c be rais$ by ! screw z desir$4 ,9 ?is is a small too!d clamp :i* c be screw$ up s z to hold a piece ( wood c>ry+ a piece ( tissue emb$d$ 9 p>aff94 ,sections c also be cut 9 celloid9 ) ?is in/rum5t1 b z oblique /rokes ) ! knife _c be made1 x is impossible to get v ?9 sections4 ,! comb9$ microtome c be obta9$ = a gu9ea f ,fraz]1 #bb ,teviot ,r[1 ,ed9burgh4 ,illu/ration3 ,fig4 #e4--;,fraz]'s ,modification ( ,ca?c>t's ,microtome4 ,a4 ,microtome arrang$ = e!r spray4 ,b4 ,cyl9d] ) clamp = hold+ object emb$d$ 9 celloid91 c4 to replace e!r spray app>atus4 ,!re is ano!r modification :i* is m g5]ally useful1 & at ! same "t m exp5sive ?an ! orig9al model4 ,9 ?is1 in/ead ( glass runn]s to support ! knife1 !re is a flat glass plate ab\t eight in*es squ>e suffici5tly l>ge to all[ ( 8,swift's pl\gh0 7fig4 #f7 be+ us$ = ! purpose ( cutt+ sections4 ,?is in/rum5t consi/s ( a triangul> brass frame1 support$ on ?ree legs1 ea* ( :i* is a screw1 tipp$ ) ivory4 ,!re is "o screw 9 front & two beh9d4 ,b5ea? ! plate1 & held 9 position by ! po/]ior screws 9 front1 & a little clamp beh9d1 is a razor ) ! edge direct$ =w>ds4 ,! edge c be rais$ or depress$ by turn+ ! ant]ior screw1 on :i* ! frame is support$4 ,be=e sections are cut ! edge ( ! razor %\ld be br\ght d[n to ! level ( ! tissue1 tak+ c>e t all ! legs are equal 9 l5g?4 ,! pl\gh %\ld !n be firmly grasp$ ) bo? h&s1 7! index f+] ( "o h& be+ left free to turn ! ant]ior screw7 & pu%$ r obliquely ?r\gh ! tissue4 ,! edge ( ! razor is !n slightly l[]$ by turn+ ! screw ?r\gh a v small angle1 & ano!r section made1 & s on4 ,) a little practice v ?9 uni=m sections may be made ) great rapidity4 ,ano!r useful e!r spray microtome is t made by ,jung ( ,heidelburg4 ,! knife sw+s r\nd a pivot1 & !re is an +5i\s rat*et arrangem5t :i* works syn*ron\sly ) ea* sw+ ( ! knife1 to raise ! tissue automatically ! requisite di/ance = ! next section to be made4 ,! exact ?ickness ( ! sections c be graduat$ ) great nicety by a simple contrivance4 ,! in/rum5t c be obta9$ 9 ?is c\ntry = ab\t #b4 ,x works satisfactorily1 b1 ) practice1 ! /ud5t w get equally good results ) ! *eap] 8,ca?c>t40 99,williams' ice freez+ microtome99 7fig4 #f74 ,?is consi/s ( a r\nd mahogany wat]-tight box provid$ ) an exit tube bel[1 & cov]$ ) a /rong plate glass lid4 ,firmly fix$ 9 ! c5tre ( ! floor ( ! box is a /\t brass pill> surm\nt$ by a brass disc :i* fits into a hole 9 ! c5tre ( ! glass lid1 s t its surface is on a level ) t ( ! lid4 ,illu/ration3 ,fig4 #f4--,williams' ,ice ,freez+ ,microtome1 ) ,swift's pl\gh4 ,to use x1 ! box is fill$ ) alt]nate lay]s ( p\nd$ ice & salt2 ! lid is !n put on & fix$ by means ( a lat]al screw4 ,! tissue to be froz5 is g5tly remov$ f ! gum & plac$ on ! brass disc & pl5ty ( gum pa9t$ r\nd x4 ,x %\ld !n be cov]$ ) a t9 cap = a few m9utes until froz54 ,sections are made ) a ,swift's ,pl\gh 7p4 #dd74 ,illu/ration3 ,fig4 #g4--,s*anze ,microtome 7see text74 99,s*anze microtome99 7fig4 #g7 is ! patt]n us$ 9 ! ,leipsic laboratories4 ,x consi/s ( a heavy iron frame ) a l>ge base4 ,! knife is c>ri$ 9 a clamp :i* slides along ! full l5g? ( ! in/rum5t1 glid+ ^u two smoo? plates ( iron :i* are arrang$ at an angle to "o ano!r4 ,! knife mu/ be mov$ v /eadily & g5tly1 z :5 us+ a long blade vibrations are easily set up :i* prev5t good sections be+ obta9$4 ,! surfaces ( contact mu/ be kept scrupul\sly free f du/1 & lubricat$ ) equal p>ts ( olive oil & ca/or oil4 ,!re are sev]al object hold]s1 :i* c be remov$ & int]*ang$1 "o connect$ ) an e!r spray app>atus1 ano!r suitable = hold+ an object emb$d$ 9 p>aff91 & a ?ird = grasp+ an object emb$d$ 9 celloid94 ,:5 celloid9 is employ$1 a specially long knife mu/ be us$1 & x mu/ be fix$ v obliquely 9 ! clamp4 ,! object hold] is rais$ by a f9e screw work$ by a l>ge brass too!d :eel4 ,!re is a rat*et arrangem5t1 by :i* ! object may be rais$ automatically any desir$ di/ance1 aft] ea* /roke ( ! knife4 ,x gives mo/ satisfactory results ) celloid9 & p>aff94 7,messrs4 ,r4 & ,j4 ,beck are ! ag5ts74 ,its co/ is ab\t #e4 99,beck]'s microtome99 is made on exactly ! same pr9ciples z ! ,s*anze4 ,! modifications are t ! c>ri] glides on glass plates in/ead ( iron ones1 & t in/ead ( ! :ole surface ( ! c>ri] be+ 9 contact ) ! plates1 !re are a few smoo? ivory buttons only4 ,friction is ?us r$uc$ to a m9imum1 & v uni=m sections c be obta9$4 ,! price is ! same z t ( ! ,s*anze4 ,fraz] has introduc$ a 8/ud5t's slid+ microtome0 on ! same pr9ciple z ! ,s*anze :i* co/s ab\t #c4 99,! ,cambridge ,rock+ microtome499--,?is in/rum5t1 z made by ! ,cambridge ,sci5tific ,in/rum5t ,company1 or ! slightly modifi$ =m made by ,messrs4 ,swift 7fig4 #h71 is ! be/ in/rum5t = cutt+ sections ( small objects emb$d$ 9 p>aff94 ,ribbons ( s]ial sections c be obta9$ f x ) great] ease & c]ta9ty ?an ) o!r microtomes4 ,?is microtome diff]s f ^? :i* h be5 previ\sly describ$ 9 t ! knife is fix$1 :ile ! object is moveable4 ,! microtome consi/s ( an oblong heavy metal /&4 ,a long b> is arrang$ s t x rides 9 see-saw fa%ion on two /rong v]tical pill>s aris+ f ! frame4 ,"o end ( ?is b> is holl[1 & receives ! piece ( wood c>ry+ ! tissue emb$d$ 9 p>aff91 :i* is firmly clamp$ 9 position4 ,?is end is depress$ by means ( a /rong spiral spr+4 ,9 ord] to raise x !re is an arrangem5t by :i* ! o!r end ( ! b> is depress$ by a cord :i* revolves r\nd a pulley4 ,:5 ! h&le is turn$1 ! tissue is rais$1 & :5 ! cord is relax$1 ! spr+ pulls ! tissue firmly & /eadily d[n4 ,! razor1 :i* mu/ h a /raight edge1 is fix$ firmly by screws1 ) its edge upw>ds at ! end ( ! microtome4 ,! object is !n adju/$ s t 9 its desc5t a ?9 slice is tak5 (f by ! razor4 ,!re is an +5i\s arrangem5t by :i* ! depression ( ! b> to raise ! section pu%es x a little fur!r 9 ! direction ( ! razor4 ,! di/ance c be graduat$ f #a_/ejj to #a_/cjjj in*4 ,! actual work+ ( ! ma*9e is !re=e v simple4 ,! position ( ! block conta9+ ! tissue to be cut hav+ be5 adju/$ s t ! razor j cuts x1 ! free end is depress$ by means ( ! pulley4 ,?is also pu%es ! section a little beyond ! razor4 ,! /rong spr+ !n draws ! tissue /eadily pa/ ! edge ( ! razor1 & a ?9 section is left on ! blade4 ,?is may be at once transf]r$ to a slide1 or if ! p>aff9 be ( ! prop] consi/5ce1 ano!r cut may be made1 :5 ! two sections %\ld adh]e by _! edges1 & s by repeat+ ! movem5t a cont9u\s ribbon may be obta9$4 ,if !re is difficulty 9 obta9+ a good ribbon1 x w usually be got ov] by tak+ a little s(t p>aff9 & atta*+ x by means ( a hot ne$le to ! l[] end ( ! p>aff9 block4 ,! co/ ( ! in/rum5t is ab\t #e4 ,illu/ration3 ,fig4 #h4--,swift's ,modification ( ! ,cambridge ,rock+ ,microtome4 99,fre% sections499--,al?\gh ^! are n s satisfactory z h>d5$ specim5s = accurate hi/ological "w1 x is (t5 v useful to make !m bo? 9 ! po/-mortem room :]e an imm$iate op9ion ( ! nature ( ! tum\r or diseas$ organ is desir$1 & also 9 ! op]at+ !atre4 ,) a little practice sections may be cut1 /a9$1 & m\nt$1 )9 t5 m9utes ( ! removal ( ! specim5 f ! body4 ,9 ?is way important in=mation may be af=d$ to ! op]at+ surgeon1 & 9 n a few cases x has caus$ ! propos$ treatm5t to be entirely alt]$4 ,?us1 9 "o case1 a suppos$ *ronic p]io/itis 0 %[n to be a s>coma1 & ! limb 0 amputat$4 ,9 ano!r1 a suppos$ s>coma ( ! ?igh 0 f\nd to be a gumma1 :5 a portion 0 remov$ & microscopically exam9$4 ,a portion ( ! specim5 %\ld be plac$ )\t any prep>ation on ! z9c plate ( ! freez+ microtome1 & "s gum pa9t$ r\nd x4 ,x is !n froz54 ,! s]um 9 ! tissues is n 9 suffici5t mass to injure ! knife :5 x is froz54 ,! knife %\ld be wett$ )1 & sections transf]r$ to1 ei!r p]ic>dial s]um1 or #c_/d p] c5t4 solution 7#gj gra9s to ! p9t71 ( common salt1 nei!r ( :i* causes ! cells to swell up z pla9 wat] does4 ,!y %\ld be c>efully float$ out on a glass slide1 an op]ation :i* requires mu* m pati5ce ?an 9 ! case ( h>d5$ sections1 z fre% sections are less coh]5t & also m /icky1 s t ! edges t5d to curl up on ! knife1 c4 ,!y %\ld !n be exam9$1 "o un/a9$1 simply m\nt$ 9 salt solution2 ano!r /a9$ ) picroc>m9e & exam9$ 9 ! sal9e solution2 & a ?ird /a9$ 9 picroc>m9e1 m\nt$ 9 ,f>rant's solution1 & pres]v$4 ,! la/ usually gives ! be/ results1 ! picroc>m9e /a9+ becom+ q brilliant aft] a week4 ,! glyc]9e1 h[ev]1 is apt to make ! sections %r9k a good deal1 & ! weight ( ! cov]-glass t5ds to break up ! unh>d5$ section4 ,,*apt] ,,iv4 ,,section ,,m\nt+4 #a4 ,by 99flotation994 ,9 ?is me?od ! section :e!r /a9$ or un/a9$ is plac$ 9 a b[l ( wat]1 or normal salt solution 7p4 #ec74 ,a clean slide is !n introduc$ into ! wat] at an angle ( ab\t #fj1 a little m ?an half ( its l5g? be+ subm]g$4 ,! section is !n br\ght up by ! ne$le & float$ z f> z possible into position on ! slide4 ,"o corn] is !n fix$ by ! ne$le1 & on g5tly )draw+ ! slide ! section %\ld lie flat4 ,if any folds are left no attempt %\ld be made to smoo!n !m out ) a ne$le1 b ! slide %\ld be re-imm]s$ until ! fold$ "p ( ! section is "u wat]4 ,x %\ld !n be g5tly )drawn1 :5 ! fold w disappe>4 ,?is manuvre mu/ be repeat$ 9 diff]5t directions until ! section lies q smoo?ly on ! slide4 ,/a9$ & un/a9$ sections are float$ out 9 ?is way be=e be+ m\nt$ 9 ,f>rant's m$ium1 & un/a9$ sections previ\s to /a9+ 9 picroc>m9e4 #b4 ,by 99transf]5ce99 ) a 99section lift]994 ,?is me?od is employ$ 9 m\nt+ 9 ,canada balsam 9 ord] to transf] ! section f ! cl>ify+ ag5t 7p4 #fc7 to ! slide4 ,! lift] is poli%$1 & ins9uat$ "u ! section4 ,! section be+ held 9 position by ! ne$le is n[ rais$ f ! fluid1 excess ( :i* is remov$ by hold+ ! section 9 position ) a m\nt$ ne$le1 & tilt+ ! lift] s z to all[ x to dra9 (f4 99,removal ( air bubbles f sections499--,:5 sections conta9 _m air bubbles1 ! be/ plan is to leave !m 9 me?ylat$ _s = a "t4 ,! bubbles !n coalesce & escape f ! section4 ,= delicate /ructures & = fre% sections ! transf]5ce to _s1 & ! subsequ5t fly+ out ( ! section :5 return$ to wat] are risky1 & ! be/ me?od ( treat+ ^! is to put ! vessel conta9+ !m "u ! receiv] ( an air pump1 if "o is available1 & slightly exhau/+ ! air4 ,! mo/ frequ5t cause ( air bubbles 9 m\nt$ specim5s1 h[ev]1 is ! employm5t ( cov]-glasses :i* h n be5 ?or\ghly cleans$4 ,prop] cleans+ is be/ effect$ by plac+ ! cov]s :5 b\ght 9 a %all[ wide m\!d /opp]$ bottle conta9+ /rong nitric acid1 & leav+ !m 9 ?is fluid = tw5ty-f\r h\rs4 ,! acid %\ld !n be dra9$ (f & wat] run ?r\gh ! vessel f a tap1 until ! wa%+s no long] give an acid reaction ) litmus pap]4 ,! wat] %\ld !n be dra9$ (f1 & ! glasses cov]$ ) absolute alcohol4 ,!y c be remov$ "o by "o & rapidly dri$ z requir$4 ,) cov]-glasses prop]ly cleans$ 9 ?is mann]1 n only w air bubbles be avoid$1 b ! cov]s w be dri$ mu* m easily ) ! clo?1 & few] w be brok5 9 ! process4 ,ano!r v frequ5t cause is ! transf]5ce ( air bubbles ) ! m\nt+ m$ium on ! glass rod4 ,?is occurs especially if ! rod be fus$ to ! /opp]4 ,! prop] bottles to use1 bo? = ,f>rant's m$ium & balsam are 8balsam bottles0 :i* h no /opp]1 b ! m\? is clos$ by a glass cap :i* fits accurately 7fig4 #i74 ,a %ort glass rod is atta*$ to ! cap1 & is us$ to transf] ! m$ium to ! slide4 ,illu/ration3 ,fig4 #i4--;,balsam bottle4 99,treatm5t ( fold$ sections499--,! fold+ may be due3-- 7#a47 ,to ! section hav+ creas$ ?r\gh be+ cut ) a knife ^: surface 0 n p]fectly smoo?4 ,?is is be/ rem$i$ by plac+ ! section 9 me?ylat$ _s = a m9ute1 & !n transf]r+ x to a b[l ( clean wat]1 :5 ! section w rapidly rise to ! top1 & spread itself out flat on ! surface ( ! wat]1 9 consequ5ce ( ! alcohol rapidly diffus+ out at ! edges into ! surr\nd+ wat]4 7#b7 ,to ! section conta9+ a l>ge am\nt ( 99fat991 z 9 ^? ( ! sk9 & subcutane\s tissue4 ,! fat may be remov$ f ! fat cells )\t mat]ially alt]+ ! appe>ance ( ! section4 ,?is is done by dehydrat+ ! section 9 alcohol1 & !n transf]r+ to a wat* glass conta9+ e!r or *lor(orm to extract ! fat4 ,! tissue %\ld be wa%$ free f e!r 9 ! alcohol & !n transf]r$ to ! b[l ( wat]1 & all[$ to float out4 ,?is process does n int]f]e ) subsequ5t /a9+ op]ations4 ,,m\nt+ ,,m$ia4 99,f>rant's solution993-- ,gum ,arabic 7pick$1 col\rless7 ,glyc]9e equal p>ts4 ,wat] ,9 mak+ ?is solution ! be/ gum arabic mu/ be us$1 & only ! cle>e/ pieces ( ?is4 8,p[d]$ gum acacia0 %\ld be avoid$1 z ?\gh x looks :ite x (t5 yields a br[n mucilage1 & besides is frequ5tly adult]at$ ) />*1 c4 ,! glyc]9e & wat] %\ld be mix$ & ! gum arabic add$4 ,! mixture %\ld be all[$ to /& = "s weeks1 ) frequ5t /irr+ until ! :ole ( ! gum is dissolv$4 ,!n all[ x to /& = a week or two long] 9 ord] t ! dirt may subside1 & ! bubbles rise to ! top4 ,! scum %\ld be remov$ & ! cle> fluid decant$ f ! s$im5t into a 8,balsam bottle0 7p4 #eh7 conta9+ a few drops ( a saturat$ solution ( ars5iate ( sodium & a small lump ( camphor4 ,if prop]ly made x is an extremely useful m\nt+ reag5t4 ,x does n cl>ify ! tissues too mu*1 & 9 consequ5ce ( its conta9+ gum x dries at ! edges & cem5ts ! cov]-glass m or less firmly 9 a week or two4 ,if ?is is n ! case ! m$ium conta9s too mu* glyc]9e & m gum mu/ be add$ to comp5sate = ?is4 ,?is dry+ at ! edge prev5ts any fur!r evaporation :ile ! glyc]9e keeps ! section p]man5tly moi/4 ,! camphor & ars5iate ( sodium prev5t ! =mation ( fungi4 ,sections pres]ve _! orig9al appe>ance 9 ?is m$ium = _m ye>s4 ,aft] a long "t !y are apt to become a little cl\dy & granul>4 ,un/a9$ sections %\ld always be m\nt$ 9 ,f>rant's m$ium1 z ! ,canada balsam process r5d]s !m q transp>5t4 ,x is suitable = almo/ any tissue /a9$ or un/a9$1 b sections ( ! n]v\s c5tres require to be m\nt$ 9 ,canada balsam1 ow+ to ! opacity ( myel9 :5 m\nt$ 9 glyc]9e4 99,canada balsam solution993--,! m$ium is made ?us3-- ,! ord9>y ,canada balsam :i* is ( a treacly consi/5ce is heat$ g5tly 9 a wat] ba? = "s h\rs1 to drive (f turp5t9e & o!r volatile oils4 ,x is !n all[$ to cool to a yell[ vitre\s mass4 ,take ( ,dri$ ,canada balsam equal p>ts4 ,xylol ,leave till dissolv$1 /irr+ occasionally4 ,unless ! solution be p]fectly cle>1 x mu/ be filt]$ ?r\gh a v ?9 pap]1 previ\sly wett$ ) xylol4 ,if ! m$ium be too ?ick m xylol %\ld be add$1 if too ?91 ! xylol %\ld be all[$ to evaporate until ! m$ium is ( ! consi/5ce ( a ?9 syrup4 ,if ! m$ium is made too ?9 mu* annoyance w be caus$ by its evaporat+ at ! edge ( ! cov]-glass1 leav+ an air-space1 :i* w increase daily until ! section is left q dry4 ,?is %\ld be rem$i$ by putt+ ano!r drop ( balsam at ! edge ( ! cov]slip & all[+ x to run 9 & displace ! air4 ,a r+ ( cem5t %\ld be put on z e>ly z possible aft]w>ds4 ,! bottle 9 :i* ! balsam is pres]v$ mu/ be v c>efully dri$ be=e be+ fill$ & !n r9s$ out ) absolute alcohol1 & aft]w>ds ) xylol4 ,turp5t9e or b5zol are (t5 us$ in/ead ( xylol 9 ! prep>ation ( ! m$ium1 & 9 ! same proportion1 b ! latt] is less apt to dissolve out ! anil9e col\rs f ! sections4 ,to m\nt sections 9 ,canada balsam !y mu/ be transf]r$ fir/ to a wat* glass conta9+ absolute alcohol or an alcoholic solution ( "s /a9+ reag5t1 e4g41 eos9e 7p4 #gb7 & left 9 x1 no attempt be+ made to spread x out1 until x is p]fectly dehydrat$1 i4e41 9 ab\t two m9utes4 ,x %\ld !n be transf]r$ to ! cl>ify+ oil on a m\nt$ ne$le1 or on a section lift]1 :i* mu/ be p]fectly dry z any spot ( moi/ure t gets on to ! section w resi/ ! cl>ify+ action ( ! oil1 & w cause unsightly opaque areas :5 ! section is m\nt$4 ,ev5 brea?+ on ! section on its way to ! cl>ify+ ag5t w prev5t uni=mity ( cle>+4 ,%\ld :ite spots appe> 9 ! section :ile 9 ! oil x mu/ be tak5 out ) z little oil z possible1 & aga9 dehydrat$ 9 absolute alcohol4 ,! process ( 99cl>ify+99 mu/ be p]=m$ 9 "s m$ium 9 :i* ,canada balsam is readily soluble1 & :i* is also readily miscible ) alcohol4 ,^? mo/ frequ5tly employ$ are oil ( cloves1 xylol1 oil ( b]gamot1 oil ( c$>1 & origanum oil4 ,! fir/ nam$ has always be5 mu* us$ because ( its agreeable od\r1 its *eapness1 & ! ease ) :i* x c be obta9$4 ,b x has ! disadvantage ( dissolv+ out _m important /a9+ reag5ts1 especially eos9e & ! v>i\s anil9e col\rs4 ,9 addition z x dissolves celloid91 sections cut 9 ?is m$ium t5d to fall to pieces :5 transf]r$ to oil ( cloves1 & "o ( ! o!r oils 7:i* h no solv5t action on celloid97 %\ld always be employ$ ) celloid9 sections4 ,oil ( b]gamot is ! mo/ g5]ally useful1 b r exp5sive4 ,:]e !re are special reasons = employ+ o!r dehydrat+ ag5ts1 !y w be indicat$ 9 ! special directions = p>ticul> /a9+ me?ods 9 ,*apt]s ,,vi4 & ,,vii4 ,z soon z ! section is plung$ into ! oil1 ! alcohol rapidly diffuses out1 s t ! edges ( ! section fly out ) x1 & ! section floats q flat on ! surface ( ! oil4 ,:5 x is completely cl>ifi$ 79 ab\t a m9ute71 z %[n by its s9k+ 9 ! oil1 x %\ld be transf]r$ to ! slide by ! section lift]1 & ! oil dra9$ (f4 ,excess ( oil may be remov$ by press+ g5tly on ! section ) a flat piece ( filt] pap] fold$ sev]al times4 ,if c>efully p]=m$ ?is manuvre w n injure ! section1 b x requires practice4 ,if ! tissue is v delicate1 & likely to be injur$ by *ang+ f "o vessel to ano!r1 or if x is l>g] ?an ! section lift] w conv5i5tly c>ry1 x %\ld be float$ out on a glass slide1 &1 z mu* wat] z possible hav+ be5 remov$ by blott+ pap]1 %\ld be dehydrat$ by add+ a little alcohol f a pipette once or twice4 ,mo/ ( ! alcohol %\ld !n be remov$ by tilt+ ! slide1 & be=e ! rema9d] has evaporat$1 "s oil ( cloves or b]gamot %\ld be add$ f ano!r pipette4 ,! section w float on ! oil at fir/1 b ! latt] w gradually come ?r\gh & appe> on ! top ( ! section4 ,:5 ?is occurs ! cl>ification is complete1 & ! oil may be run (f by tilt+ ! slide & ! section m\nt$ 9 ,canada balsam4 99,cem5t+ ( cov]-glasses499--,! cov]-glasses may be cem5t$ d[n to prev5t _! %ift+ & spoil+ ! specim54 ,if ! cov]-glass be circul>1 a ,%adbolt's turntable %\ld be us$4 ,x consi/s simply ( a horizontal heavy brass disc1 rotat+ easily on a pivot4 ,!re are a numb] ( circles trac$ on ! disc conc5trically4 ,! slide is !n fix$ on ! disc by means ( ! clips1 s t ! circumf]5ce ( ! cov]-glass corresponds to "o ( ! circles4 ,! disc is !n rotat$ & ! cem5t appli$ to ! edge ( ! cov]-glass ) a bru%4 ,_m mat]ials are employ$4 ,! mo/ suitable are3--7#a7 ,canada balsam1 :i* is almo/ col\rless & transp>5t & looks v neat4 7#b7 ,gold size4 7#c7 ,m>9e glue4 ,:5 ^! are dry a f9i%$ appe>ance may be giv5 to ! slide by lay+ on a r+ ( z9c :ite4 ,?is is made z foll[s3-- ,oxide ( z9c #a_/b dra*m4 ,b5zole half an ounce ,gum damm> ( ea*4 99,pres]vation ( sections499--,!y %\ld be kept flat1 & pres]v$ f bo? light & du/4 ,v useful c>dbo>d trays are n[ sold by almo/ all deal]s 9 boxes made to conta9 tw5ty-f\r doz5 slides = ab\t eight %ill+s1 or suitable cab9ets may be con/ruct$ by a c>p5t]4 ,,*apt] ,v4 ,,g5]al ,,/a9+ ,,me?ods4 ,mu* in=mation may be obta9$ f un/a9$ sections1 & 9 mo/ cases "o section %\ld be exam9$ un/a9$1 b ! specim5s m\nt$ 9 ?is way are s transp>5t t x is difficult to /udy ! details ( ! tissue4 ,!y are !re=e usually prep>$ by treat+ !m ) "s /a9+ reag5t1 n m]ely to r5d] !m less transp>5t1 b also to 8diff]5tiate0 ! elem5ts ( ! section1 by /a9+ "o "p m deeply ?an ano!r1 or ( a diff]5t col\r4 ,?us hmatoxyl9e /a9s ! nuclei & rapidly gr[+ p>ts ( ! tissue1 leav+ ! =m$ mat]ial1 z a rule1 mu* m lightly t9t$4 ,me?yl violet aga9 /a9s heal?y tissues blue1 & p>ts affect$ ) waxy deg5]ation a r$-violet col\r4 ,by comb9+ /a9s also mu* diff]5tiation ( ! tissue elem5ts may be obta9$4 ,sections %\ld be /a9$ ) sev]al reag5ts1 z _! effect on individual specim5s v>ies a good deal4 ,! foll[+ are ! mo/ useful /a9s = g5]al purposes3-- 99,logwood499--,?is or its purifi$ pr9ciple hmatoxyl9e is ! mo/ useful g5]al /a94 ,! hmatoxyl9e itself is pref]able1 giv+ m con/ant results1 & less diffuse /a9+4 ,= g5]al /a9+ purposes ! foll[+ =mula w be f\nd to give excell5t results3--99,hmatoxyl9e4 ,s*u*>dt's =mula499-- 7a7 ,hmatoxyl9e #c grms4 #cj grs4 ,absolute alcohol #af c4c4 #b-a_/b drms4 7b7 ,pure alum #c grms4 #cj grs4 ,di/ill$ wat] #ajj c4c4 #b ozs4 ,add 7a7 to 7b7 drop by drop & ) con/ant agitation4 ,keep = "s days expos$ to diffuse daylight until its col\r is s deep t x w n transmit ! light4 ,x %\ld !n be filt]$1 & a cry/al ( ?ymol add$4 ,x w n give v satisfactory /a9+ reactions at fir/1 & %\ld be all[$ to rip5 at lea/ a mon? or six weeks be=e us+4 ,x improves z a dye ) e mon? t x is kept4 ,:5ev] hmatoxyl9e has be5 made up ) alum z 9 ! above =mula1 an abundant r$di%-br[n precipitate =ms aft] "s "t4 ,?is 9 no way int]f]es ) ! activity ( ! solution1 b x mu/ always be filt]$ be=e be+ us$4 99,b>rett's =mula499--,introduc$ by ,dr4 ,w4 ,h4 ,b>rett1 ( ,belfa/4 ,x gives almo/ z good results z ! above4 ,x is made f ord9>y ,engli% extract ( logwood1 & is consid]ably *eap]4 ,! extract %\ld be dri$1 & f9ely p[d]$1 & !n extract$ ) absolute alcohol = sev]al days4 ,p[d]$ extract ( logwood #b grms4 #a-a_/b drms4 ,absolute alcohol #aj c4c4 #a oz4 ,filt] & add sl[ly to ,b5zoate ( sodium #a grm4 #cf grs4 ,alum #a grm4 #cf grs4 ,di/ill$ wat] #ajj c4c4 #aj ozs4 ,! /r5g? ( ! solution w v>y ) diff]5t samples ( logwood & mu/ be e/imat$ by trial4 ,?is solution is comp>atively *eap & is useful = class purposes4 99,ehrli*'s hmatoxyl9e499--,?is v useful nucle> /a9 is made z foll[s3-- 7a7 ,hmatoxyl9e #b grms4 #i grs4 ,absolute alcohol #ajj c4c4 #b ozs4 7b7 ,glyc]9e #ajj c4c4 #b ozs4 ,di/ill$ wat] #ajj c4c4 #b ozs4 ,alum #abj grms4 #b-a_/b ozs4 ,glacial acetic acid #e c4c4 #bd m9s4 ,add 7a7 sl[ly to 7b7 ) con/ant agitation4 ,all[ to rip5 9 sunlight = two mon?s be=e us+4 ,x may be employ$ z a rapid /a9 undilut$ b f> bett] results are obta9$ by us+ a weak solution1 a few drops to a wat*-glass full ( di/ill$ wat]1 & /a9+ sl[ly = f half an h\r to two h\rs4 ,! solution improves by keep+4 ,if aft] a "t ! /a9+ becomes diffuse x is an indication t ! acetic acid has evaporat$1 & a few drops m %\ld be add$4 99,kle95b]g's hmatoxyl9e499--,?is =mula diff]s f ! previ\s "o 9 be+ an alcoholic solution4 ,! calcium *loride is add$ because x 8sets up diffusion curr5ts betwe5 ! alcohol 9 ! mat]ial to be /a9$ & ! alcoholic /a9+ solution1 s enabl+ ! latt] to p5etrate m rapidly0 7,squire74 ,x is mu* us$ 9 /a9+ embryonic specim5s 9 bulk be=e emb$d+ 9 p>aff91 & 0 /rongly recomm5d$ = t purpose by ,fo/] & ,maitl& ,balf\r4 ,v>i\s =mul h be5 giv5 f "t to "t4 ,t advis$ by ,squire 7,me?ods & ,=mul1 p4 #be7 c be accurately made up )\t mu* difficulty4 7a7 ,cry/allis$ calcium *loride #bj grs4 #a_/b oz4 ,di/ill$ wat] #aj c4c4 #b drms4 7b7 ,alum #c grms4 #cb grs4 ,di/ill$ wat] #af c4c4 #agj m9s4 ,mix & add ,rectifi$ _s #bdj c4c4 #h ozs4 ,all[ x to /& & any excess ( calcium sulphate1 c41 to sep>ate4 ,filt] & add ,hmatoxyl9e #b-a_/b grms4 #be grs4 ,a little ?ymol %\ld be add$ z a pres]vative4 ,9 mak+ up ^! solutions c>e mu/ be tak5 t only di/ill$ wat] is us$1 & t all ! vessels employ$ h be5 previ\sly r9s$ out ) x1 o!rwise precipitation ( ! hmatoxyl9e w occur4 ,%\ld sections be ov]/a9$ 9 hmatoxyl9e1 ?is may be rem$i$ by wa%+ x 9 a half p] c5t4 solution ( acetic acid1 until suffici5t ( ! /a9 is dis*>g$1 b ! /a9+ is m diffuse ?an if ! happy mean _h be5 hit 9 ! fir/ in/ance4 ,hmatoxyl9e /a9s ! nuclei ( ! cells a beautiful violet col\r1 & also t9ts1 m or less lightly1 ! cell protoplasm & ! fibr\s elem5ts4 ,x also /a9s ! axis cyl9d]s ( n]ves1 & is mu* us$ 9 special /a9+ ( ! n]ve c5tres z w be describ$ lat]1 7pp4 #hh-ia74 ,! /a9 is p]man5t4 ,sections may be m\nt$ ei!r 9 ,f>rant's solution1 or 9 ,canada balsam1 ! latt] be+ pref]able4 99,eos9e499--,mu* m satisfactory results are obta9$ f ! comm]cial eos9e 7an amorph\s orange p[d] us$ 9 dye+ & 9 ! manufacture ( r$ ink71 ?an f ! pure cry/all9e =m4 ,x may be us$ z an aque\s solution 7#a_/cj p] c5t47 or z a solution 9 absolute alcohol 7#a_/ae p] c5t474 ,sections /a9$ 9 ! =m] %\ld be rapidly pass$ ?r\gh a "o p] c5t4 solution ( acetic acid 9 ord] to 8fix0 ! /a91 & !n wa%$ 9 di/ill$ wat]4 ,x is a v transp>5t /a91 & ! mo/ delicate details ( a section /a9$ ) x are p]fectly visible4 ,x /a9s ! nucleus b slightly1 :ile x /a9s ! cell protoplasm & fibr\s tissues & especially muscul> tissues a beautiful rose col\r4 ,x w be se51 !re=e1 t x /a9s ^? p>ts :i* are left un/a9$ by hmatoxyl9e1 & vice v]s4 ,?is complem5t>y action is appli$ 9 ! foll[+ me?od4 99,d\ble /a9+ ) eos9e & hmatoxyl9e499--,sections hav+ be5 /a9$ 9 hmatoxyl9e 9 ! ord9>y way1 are wa%$ 9 di/ill$ wat]1 & dehydrat$ 9 a solution 7ab\t #a 9 #aejj7 ( eos9e 9 absolute alcohol4 ,!y %\ld rema9 9 ?is = ab\t two m9utes1 & !n be pass$ ?r\gh oil ( cloves & m\nt$ 9 ,canada balsam 9 ! ord9>y way4 ,?is me?od gives extremely useful & beautiful results ) almo/ all tissues1 & is sup]ior to picroc>m9e = diff]5tiat+ ! tissue elem5ts4 ,?us1 ! nuclei are /a9$ violet1 ! cell protoplasm a mu* pal] & w>m] violet1 ! fibr\s tissues p9k1 & r$ blood corpuscles orange or brick r$4 ,! alcoholic solution ( eos9e is also us$ z a contra/ /a9 aft] /a9+ = micro-organisms ) blue or violet dyes4 99,c>m9e499--,x is made z foll[s3-- ,c>m9e 7be/7 #b #a dra*m4 ,/rong ammonia #b #a dra*m4 ,di/ill$ wat] #ajj #f ounces4 ,rub ! c>m9e ) a little wat] 9 a mort>1 add ! ammonia1 :5 ! liquid w turn black4 ,gradually add ! re/ ( ! wat]1 rubb+ x up all ! "t4 ,x %\ld be bottl$1 all[$ to /& = a few days1 & !n filt]$1 & a piece ( camphor put 9 ! bottle4 99,li?ium c>m9e99 resembles closely ammonia c>m9e 9 its /a9+ effects4 ,x is usually a matt] ( individual pref]5ce :i* is employ$4 ,c>m9e #b-a_/b grms4 #aj-a_/b grs4 ,saturat$ aque\s solution ( li?ium c>bonate #ajj c4c4 #b ozs4 ,dissolve & filt]4 ,sections may be suffici5tly /a9$ 9 ei!r ( ^! fluids 9 f ?ree to five m9utes1 b m satisfactory results are to be obta9$ by dilut+ ) tw5ty times ! bulk ( di/ill$ wat]1 & leav+ sections to /a9 = tw5ty-f\r h\rs4 ,aft] /a9+ 9 c>m9e ! sections mu/ be pass$ ?r\gh a half p] c5t4 solution ( acetic acid1 9 ord] to fix ! c>m9e 9 ! tissues1 z o!rwise ! wat] w dissolve ! /a9 out4 99,borax c>m9e99-- 7a7 ,borax #d grms4 #c dra*ms4 ,c>m9e #b grms4 #a-a_/b dra*ms4 ,di/ill$ wat] #ajj c4c4 #e ounces4 ,dissolve ) ! aid ( heat & add sl[ly to 7b74 7b7 ,alcohol #gj c4c4 #c-a_/b ounces4 ,di/ill$ wat] #cj c4c4 #a-a_/b ounce4 ,all[ to /& = a =tnight4 ,filt]1 & add a lump ( camphor4 ,to use x1 place sections1 or ! tissue 9 bulk1 9 x = f f\r to tw5ty-f\r h\rs1 accord+ to size1 & !n transf] to alcohol 7sev5ty p] c5t47 conta9+ a drop to ! ounce ( hydro*loric acid1 = tw5ty-f\r h\rs1 & !n wa% ?or\ghly 9 wat]4 ,! tissue may !n be plac$ 9 gum if x is to be froz51 dehydrat$ 9 alcohol if p>aff9 or celloid9 is to be employ$4 ,its advantage is t x is v diffusible1 & s c be us$ to /a9 tissues 9 bulk4 ,x takes a consid]able "t to /a9 suffici5tly deeply1 b !re is little fe> ( ov]/a9+4 ,x /a9s n]ve-cells & axis cyl9d]s brightly1 & also ! connective tissue1 br++ a scl]os$ pat* out v prom95tly4 99,alum c>m9e993-- ,alum five p] c5t4 solution 9 di/ill$ wat] #ajj c4c4 #a oz4 ,pure c>m9e #a grm4 #d-a_/b grs4 ,boil = tw5ty m9utes4 ,filt]4 ,add a few drops ( c>bolic acid4 ,9 us+ ?is reag5t x %\ld be filt]$ into a wat* glass1 & ! sections plac$ 9 x = at lea/ an h\r4 ,!re is no fe> ( ov]/a9+1 & !y may be left all night4 ,aft] !y h be5 /a9$ !y mu/ be ?or\ghly wa%$ 9 wat] to remove ! alum1 o!rwise num]\s cry/als ( x w be se5 9 ! field :5 ! section is m\nt$4 ,sections may be m\nt$ 9 ,f>rant's solution or 9 ,canada balsam4 ,! /a9+ effect improves v mu* aft] ! section has be5 kept a few days4 ,if desir$ its /a9+ action may be complem5t$ by dehydrat+ x 9 an alcoholic solution1 ei!r ( eos9e 7#a 9 #aejj7 or ( picric acid1 & !n cle>+ up 9 oil ( cloves1 & m\nt+ 9 ,canada balsam4 ,by itself x gives a /a9 v l t ( hmatoxyl9e1 only w>m]4 ,x picks out ! nuclei & axis cyl9d]s ( n]ves1 /a9s cell protoplasm slightly1 & ! fibr\s elem5ts sc>cely at all4 ,x may be us$ = ! same purposes z hmatoxyl9e4 ,! col\r is less attractive1 & n s deep z t ( ! latt]1 b z x does n ov]/a9 sections1 ev5 :5 left 9 x = a week1 x is a v conv5i5t /a9 = g5]al purposes4 ,x is p>ticul>ly useful z a contra/ /a9 = sections ( bra9 & sp9al cord1 aft] ! ,weig]t-,pal hmatoxyl9 process 7p4 #hh74 99,ammonia-picroc>m9e99 0 =m]ly v l>gely us$ z a /a9+ reag5t4 ,its place has n[ to a l>ge ext5t be5 tak5 by li?io-picroc>m9e4 ,9 its prep>ation ! be/ c>m9e mu/ be us$4 ,x is made z foll[s3-- ,c>m9e #a "p4 ,liq4 ammon4 =t4 #c p>ts4 ,di/ill$ wat] #c p>ts4 ,dissolve ) g5tle heat1 & add ,cold saturat$ aque\s solution ( picric acid #bjj p>ts4 ,br+ ! mixture to ! boil+ po9t1 & !n place 9 a %all[ vessel1 cov]$ ) a glass plate1 & leave x 9 full sunlight = a mon? or m4 ,filt]1 bottle1 & add six drops ( c>bolic acid to ea* ounce ( ! mixture4 ,x w keep indef9itely & improves ) age4 ,x requires filt]+ f "t to "t1 z a gelat9\s crimson mud t5ds to deposit f ! solution4 99,li?io-picroc>m9e499--,prep>$ z foll[s3-- ,c>m9e #be grms4 #aj grs4 ,saturat$ solution li?ium c>bonate #ajj c4c4 #a oz4 ,dissolve1 & add ,saturat$ solution picric acid #bej c4c4 #b-a_/b oz4 ,add a few drops ( c>bolic acid to ea* ounce4 ,x %\ld /& a day or two 9 sunlight & !n be filt]$4 ,x improves by keep+4 ,x %\ld be kept 9 a /opp]$ bottle ) a glass rod fus$ into ! /opp]4 ,:5 sections are to be /a9$ !y are to be float$ out on a clean glass slide z describ$ on page #ee4 ,! slide %\ld !n be tilt$ to all[ ! wat] to dra9 (f1 & sup]flu\s moi/ure r\nd ! section remov$ by a s(t rag1 or blott+ pap]4 ,a drop or two ( ! /a9 %\ld !n be transf]r$ to ! slide1 :i* %\ld be left ly+ q flat = ab\t t5 m9utes4 ,unless ! room is v w>m x is advisable to heat ! slide v g5tly ov] a _s lamp1 z ?is causes ! tissues to /a9 m brightly & m rapidly4 ,! excess ( ! picroc>m9e %\ld be all[$ to run (f ! slide1 & ! latt] wip$4 ,"s ( ! /a9 %\ld1 h[ev]1 be left on ! section1 z its effects g on increas+1 & are (t5 n fully se5 until a few weeks h elaps$4 ,!y %\ld be m\nt$ 9 ,f>rant's m$ium4 ,z a rule ^? m\nt$ 9 ,canada balsam d n give su* good results4 ,%\ld !re be special reasons = us+ ?is m$ium1 z 9 m\nt+ sp9al cord sections1 c41 !y %\ld be dehydrat$ aft] /a9+ 9 picroc>m9e 9 an alcoholic solution ( picric acid 7"o "p ( a saturat$ alcoholic solution to five ( alcohol71 be=e cl>ify+ 9 oil ( cloves1 z o!rwise ! alcohol w dissolve out ! picric acid1 & mu* ( ! diff]5tial /a9+ effect w be lo/4 ,! nuclei %\ld be /a9$ a bright crimson1 ! protoplasm ( ! cells yell[1 or a dull p9k1 ! fibr\s elem5ts a bright p9k1 r$ corpuscles gre51 & all dead mat]ial1 e4g41 case\s matt]1 bright yell[4 ,x also /a9s n]ve-cells1 & ! axis cyl9d]s ( n]ve fibres v brightly4 ,x is1 h[ev]1 a r unc]ta9 dye4 ,! results are mo/ brilliant 9 ! case ( fre% sections4 99,osmic acid99 is invaluable = /a9+ fatty p>ticles 9 ! cells4 ,= ord9>y use ! "o p] c5t4 /ock solution 7p4 #ba7 %\ld be dilut$ ) t5 times its bulk ( di/ill$ wat]1 & sections /a9$ 9 x all night 9 a d>k cupbo>d1 or ! wat* glass conta9+ !m may be plac$ inside a small box4 ,! sections mu/ be wa%$ ?or\ghly 9 pl5ty ( wat]4 ,if desir$ !y may be /a9$ subsequ5tly 9 picroc>m9e or me?yl violet if waxy deg5]ation also be pres5t4 ,sections %\ld be m\nt$ 9 ,f>rant's solution1 z ,canada balsam usually gives disappo9t+ results4 ,x demon/rates ! mo/ m9ute fatty p>ticles 9 deg5]at+ cells1 c41 /a9+ !m black4 ,x may be employ$ to demon/rate ! globules ( fat block+ up ! vessels 9 fat embolism4 ,x /a9s ! myel9 %ea?s ( n]ves black1 & w be aga9 ref]r$ to :5 speak+ ( me?ods ( /a9+ ! sp9al cord4 99,nitrate ( silv]99 is employ$ = /a9+ ! int]cellul> cem5t ( epi!lial cells4 ,x /a9s ?is sub/ance a deep black1 :ile ! re/ ( ! tissue takes on a br[n col\r4 ,x is us$ z a half p] c5t4 solution 9 99di/ill$99 wat]1 & kept 9 a /opp]$ bottle c>efully cov]$ up ) br[n pap]4 ,to use x take "s epi!lial tissue1 e4g41 ! om5tum f a rec5tly kill$ animal1 or a section ( "s epi!lial tum\r1 imm$iately aft] excision4 ,wa% ?or\ghly 9 di/ill$ wat] to remove all *lorides1 & !n place 9 a wat* glass conta9+ ! silv] solution4 ,keep ?is 9 ! d>k = half an h\r & !n wa% ?or\ghly 9 pl5ty ( wat]4 ,! section %\ld be m\nt$ 9 glyc]9e or ,f>rant's m$ium & kept f ! light or x w become too d>kly /a9$4 99,*loride ( gold99 is employ$ to demon/rate ! p]iph]al t]m9ations ( n]ves4 ,x c only be employ$ )9 ! fir/ half h\r aft] ! tissue has be5 remov$ f ! liv+ body4 ,! pieces ( tissue mu/ be small & may be /a9$ 9 bulk1 sections be+ subsequ5tly made4 ,a half p] c5t4 solution 9 di/ill$ wat] is employ$4 ,! tissue is transf]r$ to ?is on its removal f ! body1 until x becomes lemon col\r$4 ,x is !n expos$ 9 a "o p] c5t4 solution ( acetic acid to a /rong light until x assumes a purpli% t+e1 :i* takes f two h\rs to two days4 ,sections %\ld be m\nt$ 9 ,f>rant's m$ium4 ,x /a9s ! cells ( ! tissue1 & n]ve cells r$di% purple1 & n]ve fibrils1 especially ! t]m9al ones1 r m violet4 ,?is is v well se5 9 ! cornea4 ,x is useful sometimes = cl9ical purposes to excise a portion ( muscul> tissue & exam9e ! n]ve end+s by ?is me?od4 ,un=tunately ! /a9 is some:at unc]ta9 9 its action4 ,bett] results are obta9$ by ,sihl]'s *loral hmatoxyl9e me?od 7p4 #ib74 99,me?yl violet499--,a v satisfactory solution may be obta9$ ready made 9 ! 8telegraph5 t9te10 prep>$ by ,leonh>di1 ( ,dresd51 z recomm5d$ by ,woodhead4 ,x may also be us$ z a "o p] c5t4 solution 9 di/ill$ wat]1 a few drops ( c>bolic acid be+ add$ to prev5t ! gr[? ( fungi4 ,x is a v useful selective /a94 ,x gives two reactions1 r$ violet1 & blue violet4 ,?us x /a9s ! matrix ( hyal9e c>tilage blue violet1 b ! cells r$ violet4 ,x has also a mo/ important pa?ological application1 z x picks out any p>ts :i* h und]gone 8waxy0 or 8l>dace\s0 deg5]ation1 /a9+ !m r$ violet1 b ! re/ ( ! section blue violet4 ,ab\t t5 drops ( a "o p] c5t4 solution %\ld be filt]$ into a wat*glassful ( wat]1 & ! sections /a9$ = ab\t five m9utes4 ,!y mu/ !n be pass$ ?r\gh a half p] c5t4 solution ( acetic acid & wa%$ 99?or\ghly99 = "s "t 9 a l>ge quantity ( wat] till no m col\r comes away4 ,if ^! /eps are n tak5 ) c>e1 ! dye w diffuse out aft] ! section has be5 m\nt$1 blurr+ all details & spoil+ ! appe>ance ( ! section4 ,sections may be m\nt$ 9 ,f>rant's solution 7to :i* a spot ( =mic acid may be add$73 if m\nt$ 9 ,canada balsam ! sections mu/ be ov]/a9$ z bo? ! alcohol & oil ( cloves rapidly dissolve out ! dye4 99,safran9e499--,employ$ z a fre%ly made saturat$ solution 9 anil9e oil wat] w>m$ to #fj ,c4 7#adj ,f474 ,filt] into a wat* glass4 ,/a9 = n m ?an a m9ute4 ,dehydrate 9 alcohol :i* w remove mu* ( ! /a91 cl>ify 9 oil ( cloves or origanum oil4 ,m\nt 9 balsam4 ,ano!r me?od is to /a9 = ab\t t5 m9utes1 & !n leave = a m9ute 9 ,gram's iodide solution4 ,! sections are !n wa%$ 9 alcohol1 dehydrat$1 cl>ifi$ 9 oil ( cloves1 & m\nt$ 9 balsam4 ,by ^! me?ods ! /a9 is )drawn except f c]ta9 elem5ts1 e4g41 ^? und]go+ colloid or calc>e\s deg5]ation4 ,a qu>t] p] c5t4 wat]y solution is sometimes employ$ z x /a9s nucleoli & actively divid+ nuclei v brightly1 :ile ! re/ ( ! cell is /a9$ fa9tly4 ,x may be employ$ to /udy k>yok9esis 9 ! cells ( a rapidly gr[+ canc]4 99,ehrli*-,biondi /a9499--,?is /a9 has be5 mu* employ$ = /a9+ specim5s ( blood1 = /udy+ k>yok9esis1 & = inve/igations on ! suppos$ p>asitic bodies f\nd 9 canc] cells4 ,x is prep>$ by mix+ saturat$ aque\s solutions ( ! foll[+ anil9e dyes1 sl[ly & ) con/ant agitation3-- ,solution ( ,orange ,g #ajj p>ts ,rub9 ,s #bj ,me?yl ,gre5 ,,oo #ej f9ally add ,di/ill$ wat] #gj ,filt] f ! copi\s precipitate :i* =ms4 ,! solution mu/ be made up frequ5tly z x does n keep well4 ,sections may be /a9$ rapidly = half an h\r or an h\r1 b bett] results are obta9$ by dilut+ ! fluid ) tw5ty volumes ( wat]1 & /a9+ all night4 ,sections %\ld be wa%$ 9 wat] & !n pass$ rapidly ?r\gh absolute alcohol & xylol & m\nt$ 9 ,canada balsam4 ,,*apt] ,,vi4 ,,special ,,/a9+ ,,me?ods4--,,special ,,me?ods ,,= ,,/a9+ ,,! ,,n]ve ,,c5tres4 #a4 ,= /a9+ 99n]ve fibres994 ,?ree me?ods 7two ( :i* are modifications ( ! fir/7 are employ$ f> m (t5 ?an any o!rs4 ,by ^! me?ods ! myel9 coat+ is /a9$4 ,tissues mu/ h be5 h>d5$ previ\sly = _m weeks 9 ,mll]'s fluid1 or "s o!r bi*romate solution4 ,!y are !n ov]/a9$ 9 a solution ( hmatoxyl9e1 & ! section treat$ ) a suitable blea*+ reag5t1 :5 ! col\r is dis*>g$ f all ! tissue elem5ts except ! n]ve fibres4 ,?is me?od displays n m]ely ! n]ve fibres 9 ! :ite matt]1 b also ! f9e network 9 ! grey matt] ( ! bra9 & sp9al cord4 ,deg5]at$ fibres are left un/a9$ & s deg5]at$ tracts %ew up z un/a9$ spots on a d>k backgr\nd4 ,! sections may be subsequ5tly /a9$ ) alum c>m9e or eos9e to %ew ! cells & neuroglia4 99,weig]t's me?od499--,! piece ( cord to be cut aft] prolong$ h>d5+ 9 ,mll]'s fluid is transf]r$ )\t wa%+ to absolute alcohol & dehydrat$ prep>atory to emb$d+ 9 celloid9 7p4 #cj74 ,:5 sections are cut !y are transf]r$ at once to ,weig]t's hmatoxyl9e solution3-- ,hmatoxyl9e #a #d gra9s4 ,alcohol #aj #de m9ims4 ,di/ill$ wat] #ajj #a ounce4 ,!y are /a9$ 9 ?is = tw5ty-f\r h\rs or long]1 until !y are q black4 ,! /a9+ w take place mu* m rapidly if ! fluid be kept at #ajj ,f4 9 ! incubator4 ,aft] /a9+ !y are transf]r$ to ,weig]t's diff]5tiat+ solution3-- ,borax #b #afj grs4 ,potassium f]ricyanide #be #bjj grs4 ,di/ill$ wat] #ajj #a p9t4 ,!y are left 9 ?is solution = sev]al h\rs1 until ! gr\nd "w becomes ne>ly decol\ris$4 ,sections w sometimes /a9 m satisfactorily if !y are treat$1 accord+ to ,weig]t's orig9al directions1 = a few h\rs ) a half saturat$ solution ( acetate ( copp]4 ,if ! h>d5+ 9 ,mll]'s fluid has be5 suffici5tly prolong$1 ?is /ep is usually sup]flu\s4 99,pal's modification99 ( ,weig]t's me?od4--,by ?is me?od quick] & m complete decol\ration ( ! neuroglia1 n]ve cells1 c41 is obta9$4 ,sections are prep>$ 9 exactly ! same way z 9 ,weig]t's me?od & !n transf]r$ to ,weig]t's hmatoxyl9e4 ,pal recomm5ds t ?is solution be dilut$ to half ! /r5g? & a few drops ( a saturat$ solution ( li?ium c>bonate add$4 ,! writ] f9ds ! results equally good if ! ord9>y ,weig]t solution be employ$4 ,:5 ! sections h be5 ?or\ghly /a9$ !y are wa%$ 9 di/ill$ wat] & plac$ 9 a ?ree-qu>t] p] c5t4 solution ( p]manganate ( potassium4 ,! "t requir$ 9 ?is solution dep5ds on ! "t ! specim5 has be5 9 ,mll]'s fluid4 ,x %\ld n be less ?an half a m9ute1 & 9 v ?or\ghly h>d5$ specim5s1 five m9utes may be all[$ ) advantage4 ,9 ?is solution ! sections w become ( an opaque br[n col\r4 ,!y are wa%$ 9 di/ill$ wat]1 & transf]r$ to3-- 99,pal's diff]5tiat+ solution499 ,potassium sulphite #a grm4 #dj gr4 ,oxalic acid #a grm4 #dj gr4 ,di/ill$ wat] #bjj cc4 #a p9t4 ,!y are kept 9 ?is = "o to five m9utes1 accord+ to ! dep? ( /a9+1 until ! :ite & grey matt] are cle>ly def9$ & ! br[n col\r is completely dis*>g$4 ,if ! br[n /a9 does n readily cle> up1 ! section %\ld be return$ to ! p]manganate solution = ab\t half a m9ute1 & aga9 treat$ ) 8,pal's solution40 ,?is manuvre may be repeat$ sev]al times4 ,z soon z ! sections are ?or\ghly diff]5tiat$ !y are transf]r$ "o by "o to a l>ge vessel ( wat] & ?or\ghly wa%$4 ,! blue /a9 ( ! hmatoxyl9e becomes bright] dur+ ! wa%+ process4 ,! sections may be m\nt$ at once1 b m beautiful results w be obta9$ if !y are /a9$ 9 alum c>m9e = #bd h\rs4 ,!y %\ld !n be wa%$1 dehydrat$ 9 alcohol1 cl>ifi$ 9 oil ( b]gamot1 & m\nt$ 9 ,canada balsam4 ,! v prolong$ h>d5+ 9 ,mll]'s fluid :i* is a necess>y prelim9>y = ?is me?od l$ to ! introduction (3-- 99,s*f]'s modification99 ( ,pal's me?od4--,9 ?is me?od h>d5+ 9 ,mll]'s fluid = ?ree or f\r weeks is suffici5t4 ,! sections are made exactly z 9 ! previ\s me?od1 & transf]r$ to ,m>*i's fluid 7p4 #bd7 = six h\rs4 ,!y are wa%$ & /a9$ all night 9 ! foll[+3-- ,hmatoxyl9 #a #d grs4 ,alcohol #aj #de m94 ,acetic acid 7#b p] c5t4 aque\s solution7 #ajj #a oz4 ,! subsequ5t processes ( diff]5tiation1 blea*+1 c41 are exactly ! same z 9 ,pal's me?od4 99,osmic acid499--,employ$ ) fre% & also ) h>d5$ specim5s to demon/rate ! m$ull>y %ea?4 ,mu* ! be/ results are obta9$ ) ! =m]4 ,! n]ves1 or small pieces ( ! c5tral n]v\s sy/em are plac$ 9 half to "o p] c5t4 solution ( osmic acid z soon z possible aft] dea? & kept 9 ! d>k = ab\t a week4 ,! tissue mu/ be v ?or\ghly wa%$ 9 runn+ wat] to remove all traces ( osmic acid1 & !n /a9$ = a c\ple ( days 9 borax c>m9e to demon/rate ! nuclei & axis cyl9d]s4 ,sections may be made 9 gum1 or ! tissue may be teas$ ) ne$les & !n m\nt$ 9 ,f>rant4 ,emb$d+ 9 celloid91 & m\nt+ 9 balsam are inadvisable1 because ! e!r t5ds to dissolve out myel91 & ! cl>ify+ oil to r5d] x too transp>5t4 #b4 99,intra-muscul> ramifications ( n]ves993-- 99,sihl]'s *loral hmatoxyl9e me?od499--,?is me?od reveals ! intra-muscul> n]ve-end+s1 & also br+s into prom95ce ! curi\s 8muscle sp9dles0 :i* ,%]r+ton has %[n to be connect$ ) ! po/]ior n]ve roots1 & :i* are believ$ by "s to be ! end organs subs]v+ muscul> s5se4 ,a piece ( muscle is tak5 z soon z possible aft] dea?1 or f an amputat$ limb1 & slices cut ab\t "o-t5? ( an in* ?ick ) ! freez+ microtome4 ,transf] = tw5ty-f\r h\rs to ! foll[+ solution3-- ,acetic acid #a "p4 ,glyc]9e #a ,"o p] c5t4 aque\s solution ( *loral hydrate #f p>ts4 ,! tissues swell up 9 ?is fluid & become transluc5t & gelat9\s 9 appe>ance4 ,!y are n[ plac$ 9 pure glyc]9e until saturat$ z %[n by _! s9k+ to ! bottom ( ! di%4 ,?is usually takes sev]al days4 ,!y may n[ be /a9$ 9 ! foll[+ solution3-- ,ehrli*'s hmatoxyl9e 7p4 #gj7 #a "p4 ,glyc]9e #a ,"o p] c5t4 aque\s solution ( *loral hydrate #f p>ts4 ,!y may be left 9 ?is f ?ree days to a week ) little fe> ( ov]/a9+4 ,portions may !n be teas$ ) ne$les1 & m\nt$ 9 glyc]9e1 or ! /a9$ tissue may be press$ out into a suffici5tly ?9 lay] by squeez+ x =cibly betwe5 two glass slides4 99,motor & s5sory n]ve end+s499--,^! are be/ /a9$ by ! *loride ( gold me?od 7p4 #hb74 ,specim5s mu/ be tak5 f ! body imm$iately aft] dea?4 ,! me?od is !re=e useless = ! po/-mortem room1 b may be us$ = tissues remov$ by op]ation4 ,small pieces ( tissue mu/ be employ$ & mu/ be /a9$ 9 bulk1 sections be+ made subsequ5tly4 ,= motor n]ve end+s ! muscle ( a frog or human muscle f a limb j amputat$ may be tak54 ,specim5s %\ld be prep>$ aft] /a9+ by teas+ 9 pref]5ce to mak+ sections4 ,m\nt 9 ,f>rant4 ,s5sory n]ve end+s may be conv5i5tly /udi$ 9 ! cornea ( a rec5tly kill$ frog or rabbit1 or 9 a fre%ly extirpat$ human eye4 ,tactile end organs may be /udi$ 9 ! lip or f+] tips1 ta/e buds 9 ! papilla foliata ( ! rabbit's tongue1 & ,pac9i's corpuscles are well se5 9 ! mes5t]y ( a ?9 cat4 #c4 99,/a9+ n]ve cells499 99,bevan ,lewis's anil9e blue-black me?od993--,?is me?od is ! be/ = demon/rat+ ! weal? ( n]ve cells 9 ! fre% c]ebral cortex4 ,! solution ( anil9e blue-black %\ld be ( ! /r5g? ( #a 9 #djj1 ab\t a gra9 to ! ounce4 ,a piece ( ! c]ebral cortex ) pia mat] atta*$1 %\ld be remov$ z soon z possible aft] dea? by p>allel cuts ab\t #a_/h in* ap>t1 & p]p5dicul> to ! surface ( ! convolution1 plac$ on ! plate ( ! freez+ microtome & j froz5--n too h>d or ! tissue w be brittle & w also injure ! edge ( ! razor4 ,z soon z a good section is obta9$ ! razor %\ld be plung$ into a l>ge b[l ( cold wat] to deta* ! section1 :i* is at once float$ on a glass slide1 & osmic acid solution1 #a_/d p] c5t4 all[$ to fl[ ov] x f a pipette4 ,?is w fix ! tissue elem5ts 9 ab\t two m9utes4 ,! section is aga9 float$ (f into ! b[l ( wat] & ?or\ghly wa%$ to free x f ! osmic acid4 ,x is !n /a9$ ei!r on ! slide1 or 9 a wat*-glass1 ) ! anil9e blue-black solution = an h\r 9 ! cold1 or half-an-h\r if ! solution is slightly w>m$4 ,! dye is ?or\ghly wa%$ away ) di/ill$ wat]1 excess ( moi/ure wip$ (f ! slide ) blott+ pap]1 & ! section all[$ to dry "u a glass bell j>4 ,x is n practicable to dehydrate by means ( alcohol z x w\ld cause sudd5 %r9k+ ( ! tissues4 ,:5 ! section is dry a drop ( ,canada balsam is appli$ & x is cov]$ ) alcohol 9 ! usual way4 ,! n]ve cells & _! processes are /a9$ a deep slate col\r1 z are ! nuclei ( ! connective tissue cells1 :ile ! gr\nd "w ( ! neuroglia is fa9tly /a9$ & ( a neutral grey t9t4 ,?is me?od gives beautiful results bo? ) normal & morbid specim5s4 ,v>i\s o!r anil9e dyes1 indul91 me?yl5e blue1 g5tian violet1 h be5 employ$ 9 ! same way1 b none ( !m give su* good or uni=m results z anil9e blue-black4 ,h>d5$ specim5s may also be /a9$ ) anil9e blue-black1 b ! results are n to be comp>$ ) ^? obta9$ by ! fre% me?od4 ,! /a9 is usually diffuse1 b ?is c be improv$ by plac+ ! sections = #a_/b - #b m9utes 9 a #b p] c5t4 solution ( *loral hydrate 9 di/ill$ wat] aft] /a9+4 99,golgi's metallic /a9s99 = n]ve cells4 ,golgi introduc$ ! me?ods ( produc+ a metallic deposit ( m]cury or silv] 9 ! n]ve cell1 reveal+ bo? ! cell & its processes4 ,?is me?od has be5 v fruitful 9 discov]ies1 especially 9 ! h&s ( ,ramon y ,cajal1 ,klli*]1 ,van ,gehuct5 & o!rs4 ,x gives be/ results ) embryonic tissues4 ,to ensure good results x is important t ! tissue be remov$ imm$iately aft] dea?4 ,sections ( bra9s remov$ "s h\rs aft] dea? usually give disappo9t+ results4 ,!re are sev]al me?ods n[ 9 vogue1 all slight modifications ( ,golgi's orig9al me?ods4 99,silv] nitrate me?od499--,small pieces n m ?an a qu>t] ( an in* cube1 are transf]r$ /raight f ! body to a l>ge quantity ( ,m>*i's fluid 7p4 #bd7 & kept 9 x = ab\t a week1 or long] 9 ! case ( adult specim5s4 ,on removal f ,m>*i's solution ! tissue %\ld be wa%$ = a few seconds 9 di/ill$ wat]1 & !n plac$ 9 a l>ge quantity ( a #c_/d p] c5t4 solution ( nitrate ( silv] solution 9 di/ill$ wat] = at lea/ a week4 ,! lump ( tissue becomes ( a brick r$ col\r ow+ to a coat+ ( silv] *romate4 ,on removal f ! silv] solution ! tissue %\ld be wa%$ 9 me?ylat$ _s = a few m9utes & ! incru/ation ( silv] *romate bru%$ (f4 ,sections may be cut 9 gum & celloid92 or !y may be fix$ on a cork ) celloid9 or _s v>ni% & cut )\t emb$d+3 v ?9 sections are n requir$4 ,dehydrate 9 alcohol1 cle> 9 xylol1 & m\nt 9 balsam4 ,goodall advises a mixture ( pyrid9e & xylol = cle>+1 & m\nts 9 /rong xylol-damm> solution1 )\t a cov]-glass4 ,v c>eful att5tion to details & mu* practice are requir$ be=e uni=mly good results c be obta9$4 ,! results are extremely beautiful & well repay ! lab\r exp5d$ on !m4 ,! cells & _! processes appe> black on a yell[i% gr\nd4 ,a me?od has be5 employ$ = deep5+ ! col\r ( ! /a91 b ! writ] has no exp]i5ce ( x4 ,kallus 7,zeits*4 f4 ,wiss4 ,mikr41 #ahic1 #dgg7 dilutes an ord9>y hydrok9one develop+ solution 7prep>$ z = develop+ an ord9>y photographic plate7 ) ab\t t5 times its volume ( di/ill$ wat]4 ,j be=e us+ a ?ird "p ( absolute alcohol is add$4 ,sections :i* h be5 ?r\gh ! silv] process :5 plac$ 9 x become grey or black 9 a few m9utes1 &1 aft] wa%+ 9 me?ylat$ _s1 are transf]r$ to a #bj p] c5t4 aque\s solution ( hyposulphite ( soda = a c\ple ( m9utes & !n wa%$ v ?or\ghly 9 di/ill$ wat] = tw5ty-f\r h\rs4 ,!y are !n dehydrat$ & m\nt$ 9 balsam4 99,buckley's modification ( ! silv] me?od499--,describ$ 9 ,bra91 ,w9t] numb]1 #ahie4 ,! me?od is applicable to specim5s t h be5 h>d5$ 9 ,mll]'s fluid4 ,?9 slices are cut 9 ! usual way1 & !n imm]s$ 9 ,bi*romate ( potassium1 #c p] c5t4 solution #e p>ts ,osmic acid1 #a p] c5t4 solution #a "p = ?ree to five days4 ,excess ( bi*romate is remov$ f ! sections by blott+ pap]1 & !y are transf]r$ to ! fre%ly prep>$ /a9+ mixture3-- ,phospho-molybdic acid 7#aj p] c5t47 #a m9im #b drops4 ,nitrate ( silv] 7#a p] c5t47 #a ounce #fj c4c4 :i* mu/ n be filt]$4 ,/a9 = sev]al days4 ,! sections %\ld be cut at once aft] removal f ! /a9+ solution4 ,x is claim$ t ! m9ute details ( /ructure ( ! cell processes are bett] %ewn by ?is me?od4 99,corrosive sublimate99 me?od4--,?is me?od is simil> 9 its mode ( action to ! la/1 m]cury be+ deposit$ 9 ! cell in/ead ( silv]4 ,x is r less c]ta9 & requires m practice4 ,x seldom /a9s uni=mly4 ,"o cell w be f\nd exquisitely /a9$ :ile ^? 9 its vic9ity are unaffect$4 ,small pieces ( cortex are h>d5$ = sev]al weeks 9 ,mll]'s fluid1 or o!r bi*romate solution1 & are !n transf]r$ direct to a "o-half p] c5t4 aque\s solution ( corrosive sublimate1 9 :i* !y %\ld be left f ?ree to six weeks4 ,%ort] p]iods w only give disappo9t+ & incon/ant results4 ,sections %\ld be cut1 if possible1 9 gum4 ,!y may be m\nt$ 9 ,f>rant1 or dehydrat$ & m\nt$ 9 balsam4 ,tal has propos$ to r5d] ! effect %>p] by trans=m+ ! deposit ( m]cury into m]curic sulphide1 by treat+ ! sections ) a solution ( sulphide ( sodium1 :i* he prep>es by saturat+ a t5 p] c5t4 solution ( cau/ic soda ) sulphurett$ hydrog5 & !n add+ an equal quantity ( fre% soda solution4 ,!y are /a9$ 9 ?is = a few m9utes & !n ?or\ghly wa%$4 ,by ?is me?od ! pyramidal cells & _! delicate processes appe> z black opaque objects on a light gr\nd4 ,! neuroglia cells ) _! f9e delicate processes are (t5 also beautifully /a9$4 99,nissl's anil9e me?od499--,?is me?od is complem5t>y to ,golgi's me?od4 ,! latt] impregnates ! cell r5d]+ x opaque & %ew+ its =m ) great def9it5ess4 ,nissl's me?od /a9s ! protoplasm )\t greatly r$uc+ its transp>5cy & all[s u to /udy details ( cell /ructure4 ,small portions ( tissue1 remov$ z soon z possible aft] dea?1 are h>d5$ 9 alcohol4 ,sections are !n cut1 pref]ably 9 gum1 z celloid9 is inconv5i5t ow+ to its /a9+ s deeply ) anil9e dyes4 ,sections are transf]r$ f alcohol to a "o-half p] c5t4 aque\s solution ( me?yl5e blue1 :i* is heat$ 9 a wat* glass till x /eams freely1 b %ort ( ! boil+ po9t4 ,/a9 = ab\t a qu>t] ( an h\r & all[ to cool4 ,transf] ! sections to a mixture conta9+ "o "p ( anil9e oil & t5 ( absolute alcohol1 & move !m ab\t till no m col\r comes away4 ,transf] ! section to a slide ) a section lift]1 dra91 & dry well by press+ fold$ filt] pap] c>efully on ! section4 ,all[ "s origanum oil to fl[ ov] ! section & remove excess ( ?is by pressure ) blott+ pap]4 ,moi/5 ) b5z9e1#a & add a drop ( colophonium res9 dissolv$ 9 b5z9e4 ,! slide is w>m$ cauti\sly till ! b5z9e is driv5 (f & ! colophonium liquefi$ by heat alone1 & !n ! cov]-glass is appli$4 #a ,! /ud5t w be> 9 m9d ! dang] ( work+ ) b5z9e ne> a nak$ light4 ,mag5ta & o!r anil9e dyes may also be employ$ 9 a simil> mann]4 ,,*apt] ,,vii4 ,,special ,,me?ods ,,= ,,/a9+ ,,micro-,,organisms ,,& ,,blood4 ,x is impossible1 )9 ! limits ( ?is "w1 to attempt any adequate description ( ! mod]n me?ods ( bact]iological inve/igation4 ,"s ( ^! are v l5g?y & complicat$1 & require mu* skill & practice be=e good results c be reli$ on4 ,b ^? :o d n desire to make a special /udy ( bact]iology may (t5 require to exam9e = ! pres5ce ( organisms 9 sections1 or 9 v>i\s excretions1 & x is hop$ t !y may f9d ! foll[+ %ort description ( special me?ods suffici5t = _! purpose4 ,= m elaborate "w !y mu/ consult "o ( ! _m excell5t textbooks on ! subject4 ,! /ud5t %\ld provide himself ) ! foll[+ dyes 9 p[d]3-- 99,me?yl5e blue499 99,g5tian violet499 99,me?yl violet499 99,fu*s9e499 99,bism>ck br[n499 ,! foll[+ solutions ( ^! dyes are us$3-- #a4 ,saturat$ alcoholic solutions :i* may be kept 9 /opp]$ bottles4 #b4 ,"o p] c5t4 aque\s solutions4 ,^! mu/ be fre%ly made ea* "t ( us+4 ,9 filt]+ ei!r alcoholic or aque\s solutions x is well to moi/5 ! filt] pap] be=eh& ) alcohol or wat] z ! case may be4 ,! foll[+ special solutions w also be want$3-- 99,lffl]'s me?yl5e blue499--,9 ?is solution a weak solution ( cau/ic pota% is employ$ z a mordant3-- ,saturat$ alcoholic solution ( me?yl5e blue #c volumes4 ,cau/ic pota%1 aque\s solution #a 3 #aj1jjj #aj volumes4 ,filt]4 ,?is solution is p]haps ! mo/ g5]ally useful /a94 ,x col\rs mo/ bacilli & micrococci1 & :ile rapid 9 its action r>ely ov]/a9s4 ,x mu/ be made up fre% on ea* occasion4 ,x is ! be/ c\nt]/a9 aft] /a9+ tub]cle bacilli1 c41 ) fu*s9e4 99,ziehl's c>bol-fu*s9e499 ,c>bolic acid 7#e p] c5t4 aque\s solution7 #ajj volumes4 ,fu*s9e 7saturat$ alcoholic solution7 #aa volumes4 ,! solution mu/ be filt]$ imm$iately be=e be+ us$4 99,gram's iod9e solution499--,sections are plac$ 9 ?is solution aft] be+ /a9$ ) anil9e dyes4 ,! iod9e 9 "s way fixes ! dye 9 ! organisms1 s t !y are n decol\ris$ along ) ! re/ ( ! tissues4 ,x is made ?us3-- ,iod9e #a grm4 #a-a_/b gra9s4 ,iodide ( potassium #b grms4 #c gra9s4 ,di/ill$ wat] #cjj c4c4 #a ounce4 ,t5 p] c5t4 aque\s solutions ( 99nitric99 & 99sulphuric99 acids %\ld be prep>$ & may be kept indef9itely4 ,! foll[+ are ! g5]al me?ods ( employ+ ^! reag5ts = ! purpose ( /a9+ organisms 9 sections4 ,special me?ods are requir$ = special organisms1 b "o or two only c be giv54 99,weig]t's me?od499--,! sections mu/ be plac$ 9 a fre%ly made "o p] c5t4 aque\s solution ( me?yl violet1 g5tian violet1 fu*s9e1 c4 ,! solution may be kept at ! temp]ature ( ! body 9 an incubator4 ,! organisms w (t5 /a9 m readily if ! section be pass$ ?r\gh a #a 9 #bjjj solution ( corrosive sublimate be=e putt+ x into ! /a9+ fluid4 ,aft] /a9+ ! section is wa%$ 9 di/ill$ wat] & !n 9 me?ylat$ _s until x appe>s almo/ decol\ris$4 ,"s pref] to decol\rise ! tissues by wa%+ 9 a half p] c5t4 solution ( acetic acid in/ead ( me?ylat$ _s4 ,practice is requir$ be=e ! correct "t = decol\ris+ is accurately e/imat$4 ,! beg9n] %\ld float a section rapidly on ! slide n[ & !n1 put on a cov]-glass & exam9e x "u a l[ p[] to see if ! decoloration has be5 c>ri$ f> en\gh4 ,a contra/ /a9 may !n be us$1 su* z picroc>m9e1 aft] :i* ! section may be m\nt$ 9 ,f>rant's m$ium3 or a weak solution ( ano!r anil9e col\r may be us$ z a c\nt] /a91 aft] :i* ! section is cl>ifi$ 9 xylol1 & m\nt$ 9 balsam dissolv$ 9 xylol4 99,gram's me?od499--,place "s anil9e oil 9 a te/ tube & add t5 times its volume ( di/ill$ wat]4 ,close ! end ) ! ?umb & %ake v ?or\ghly4 ,filt] n9ety drops into ano!r clean te/ tube1 & add t5 drops ( a saturat$ solution ( g5tian violet or "s simil> dye4 ,filt] ! mixture into a wat* glass4 ,/a9 sections 9 x = f ?ree m9utes to half-an-h\r accord+ to ! temp]ature1--! %ort] "t = ! incubator at #ajj1 ! long] :5 ! sections are /a9$ at ! ord9>y temp]ature ( ! room4 ,wa% 9 di/ill$ wat]1 & transf] to ,gram's iod9e solution until !y become black1 usually 9 a few m9utes4 ,!y are !n decol\ris$ 9 absolute alcohol4 ,?is (t5 takes "s "t4 ,x may be ha/5$1 z ,crook%ank sugge/s1 by plac+ ! section 9 clove oil1 return+ to alcohol1 & s on4 99,ehrli*'s modification99 ( ,gram's me?od4 ,! contra/ /a9 is "h us$ fir/4 ,/a9 ! section 7e4g41 t ( a mitral valve 9 a case ( ulc]ative endoc>ditis71 9 an alcoholic solution ( eos9e 7#a 9 #aejj74 ,transf] to a solution ( "s anil9e dye1 su* z g5tian violet1 dissolv$ 9 anil9e oil wat]1 exactly z 9 ,gram's me?od4 ,! section floats on ! surface & spreads out1 ow+ to ! alcohol diffus+ out4 ,/a9 = ab\t tw5ty m9utes4 ,wa% ! section 9 wat]1 & float out 7p4 #ee7 on a glass slide4 ,all[ ! wat] to dra9 (f & add ,gram's iod9e sl[ly f a pipette s z n to dis>range ! section4 ,:5 ! section has become q black p\r (f ! ,gram's solution4 ,remove all sup]flu\s fluid f ! slide ) blott+ pap]1 & dry ! section by c>efully & firmly press+ on x a fold$ piece ( blott+ pap]4 ,if ?is is done ) c>e ! section ne$ n be injur$ 9 ! lea/4 ,decoloration is effect$ on ! slide ) anil9e oil1 in/ead ( alcohol z 9 ! prec$+ me?od4 ,! slide is rock$ ab\t s t ! col\r may be ev5ly dis*>g$ by ! anil9e4 ,:5 no m col\r comes away1 ! anil9e oil is p\r$ (f1 ! section cl>ifi$ 9 xylol1 & m\nt$ 9 ,canada balsam4 ,z soon z ! section is decol\ris$ x may be treat$ ) a contra/ /a91 ! mo/ suitable be+ alcoholic solutions ( eos9e or ,bism>ck br[n if a blue /a9 has be5 employ$1 or me?yl5e blue if fu*s9e has be5 ! fir/ /a9 us$4 ,! foll[+ w be f\nd ! mo/ useful /a9s & contra/ /a9s3-- ,,/a9s4 ,,contra/ ,,/a9s4 ,picroc>m9e4 ,g5tian violet4 ,eos9e4 ,me?yl violet4 ,bism>ck br[n4 ,me?yl5e blue4 ,safran9e4 ,mag5ta4 ,me?yl5e blue4 ,fu*s9e4 ,& vice v]s4 ,mu* practice is requir$ 9 us+ ei!r ( ! me?ods be=e "o c judge accurately h[ long to leave sections 9 ! /a9+ reag5ts or decol\ris+ ag5ts1 & ! beg9n] mu/ n be disc\rag$ if at fir/ he is unable to obta9 good results al?\gh he foll[s ! book directions mo/ m9utely4 99,ehrli* me?od = tub]cle bacilli499--,sections are /a9$ = six to tw5ty-f\r h\rs 9 a "o p] c5t4 solution ( g5tian violet1 me?yl violet1 me?yl blue or fu*s9e4 ,!y w /a9 m rapidly if ! /a9+ fluid be kept 9 an incubator at ! body temp]ature4 ,!y %\ld be remov$ f ! /a9+ fluid1 & wa%$ 9 di/ill$ wat]1 & !n transf]r$ 7pref]ably on a glass section lift]7 to a t5 p] c5t4 solution ( nitric acid 9 di/ill$ wat] until !y are ne>ly decol\ris$4 ,!y %\ld !n be v ?or\ghly wa%$ 9 di/ill$ wat]4 ,!y may !n be treat$ ) "s suitable contra/ /a9 & m\nt$ 9 ,canada balsam4 99,neels5's /a9 = tub]cle bacilli499--,sections are plac$ 9 ,ziehl's c>bol-fu*s9e solution 7p4 #ajc7 :i* %\ld be w>m$ = t5 m9utes to half-an-h\r4 ,!y are !n decol\ris$ 9 a solution ( sulphuric acid4 ,tw5ty-five p] c5t4 is ! /r5g? orig9ally recomm5d$1 b a t5 p] c5t4 solution does equally well & injures ! section less4 ,!y are !n v ?or\ghly wa%$ 9 a l>ge quantity ( wat]1 & aft]w>ds may be treat$ ) a contra/ /a94 99,gibbes' d\ble /a9 = tub]cle bacilli499-- 7#a7 ,rosanil9e hydro*lorate #b grms4 #be grs4 ,me?yl blue #a grm4 #abe grs4 ,triturate 9 a glass mort>1 7#b7 ,anil9e oil #c c4c4 #cge grs4 ,rectifi$ _s #ae c4c4 #c-a_/b drms4 ,dissolve & add sl[ly to 7#a74 7#c7 ,la/ly add sl[ly to ! mixture ,di/ill$ wat] #ae c4c4 #c-a_/b drms4 ,"s ( ! solution is filt]$ into a wat* glass & w>m$4 ,! sections are plac$ 9 x & left = "s h\rs4 ,!y are !n wa%$ 9 me?ylat$ _s till !y are suffici5tly decol\ris$1 & !n rapidly pass$ ?r\gh absolute alcohol & oil ( cloves & m\nt$ 9 balsam & xylol4 ,x is a v useful /a9 = exam9+ ! sputum = tub]cle bacilli4 ,9 ord] to /a9 fluids1 su* z blood1 pus1 or sputum1 = organisms1 a v ?9 lay] %\ld be obta9$ by plac+ a little ( ! fluid betwe5 two clean cov]-glasses & press+ !m toge!r4 ,!y are !n sep>at$ & all[$ to dry4 ,! film is fix$ by hold+ ! cov]-glass 9 a pair ( =ceps1 & pass+ x sl[ly ?r\gh ! flame ( a _s lamp two or ?ree times4 ,films ( pus %\ld be #8cle>$' aft] fix+ by plac+ !m 9 a tw5ty p] c5t4 solution ( acetic acid = ?ree m9utes4 ,= cl9ical purposes x is (t5 necess>y to exam9e ur9e1 fces or vomit$ matt] = bacilli4 ,films are prep>$ 9 ! usual way & all[$ to evaporate sl[ly1 & !n fix$ by pass+ ?r\gh ! flame1 & !n wa%$ 9 di/ill$ wat] be=e /a9+4 ,9 ! case ( vomit$ matt] & fces ?is is usually done )\t difficulty4 ,9 ! case ( ur9e h[ev] x is (t5 difficult to get ! ur9e to evaporate completely4 ,a syrupy lay] rema9s1 & if m heat be appli$ x decomposes & *>s1 & ! products cause precipitation ( anil9e dur+ subsequ5t /a9+ processes4 ,?is may be p>tly avoid$ by g5tly wa%+ ! film 9 di/ill$ wat] be=e /a9+4 ,ano!r plan is to mix ! ur9>y deposit ) a little gelat9e free f organisms1 su* z t 9 unus$ culture tubes4 ,! gelat9 is liquefi$ by heat1 & mix$ ) ! deposit4 ,films are made f ?is mixture1 & all[$ to set1 & !n ?or\ghly wa%$ 9 di/ill$ wat]4 ,! film is !n dri$ ?or\ghly1 & ! cov]-glass laid flat ) ! film upp]mo/1 & a few drops ( ! /a9+ fluid filt]$ on to x4 ,aft] x has be5 /a9$ suffici5tly ! /a9 is dra9$ (f1 & ! slip g5tly wa%$4 ,! film may !n be /a9$ ) "s contra/ /a9 9 exactly ! same way z sections1 aga9 wa%$1 dri$ betwe5 folds ( blott+ pap]1 & m\nt$ 9 balsam4 ,x is sometimes difficult to tell :i* is ! side ( ! cov]-glass :i* be>s ! film4 ,?is is readily done by hold+ ! glass obliquely s t light f a w9d[ is reflect$ f its surface4 ,! side :i* is coat$ appe>s dull2 :ile ! o!r is smoo? & bright4 ,,me?ods ,,( ,,exam9+ ,,blood4 ,9 all ^! me?ods blood is obta9$ by prick+ ! sk9 ( "o ( ! f+]s1 or ! lobule ( ! e>1 pref]ably ! latt]4 ,! sk9 mu/ previ\sly be wa%$ ) soap & wat] or e!r1 to remove any grease or epi!lial scales4 ,! puncture %\ld be made firmly s t blood may escape freely4 ,! f+] or e> mu/ n be squeez$4 ,specim5s mu/ be made rapidly be=e r$ corpuscles h run into r\leaux4 ,! slides & cov]slips employ$ mu/ be scrupul\sly clean1 or x is impossible to get really good films4 ,!y %\ld be clean$ ) nitric acid & alcohol accord+ to ! directions on page #eg4 ,fre% specim5s %\ld be exam9$4 ,! cov]slip is made j to t\* ! drop ( blood at "o edge1 s z to transf] a small quantity only1 & is at once l[]$ on to ! slide ) ! aid ( a m\nt$ ne$le4 ,if slide & cov]slip be p]fectly clean ! blood w spread out into a ?9 film1 ! corpuscles ly+ q flat4 ,if !re be any delay1 or if ! cov]-glass be n q clean ! r$ corpuscles w run into masses & ! specim5 w be useless = m9ute exam9ation4 ,ano!r specim5 may be mix$ ) a little ( ,f]ri]'s solution 7p4 #abi7 be=e m\nt+4 ,p]man5t cov]slip films may also be prep>$4 ,"h aga9 ! use ( absolutely clean cov]slips is ess5tial1 & ! blood mu/ be tak5 imm$iately x escapes f ! puncture4 ,a little blood is tak5 on a cov]-glass :i* is held horizontally4 ,ano!r cov]-glass is l[]$ on to ?is & by its weight & by capill>y attraction1 ! drop ( blood quickly becomes trans=m$ into a ?9 film4 ,! two cov]s are sep>at$ z soon z ! film is =m$ by rapidly slid+ !m (f "o ano!r4 ,?is manuvre requires a little practice & dext]ity4 ,! movem5t ( ! slips mu/ be 9 an exactly p>allel direction o!rwise ! coat+ left w be unev51 j z :5 two pieces ( bread & butt] are pull$ ap>t4 ,ev5 ) practice x is difficult to get m ?an "o good film1 ! l[] be+ usually be/4 ,!re are f\r ways ( fix+ ! film4 #a4 ,exposure to 99osmic acid vap\r994 ,! film :ile /ill moi/ is held ov] ! m\? ( a bottle conta9+ at lea/ "o p] c5t4 solution ( osmic acid4 ,9 a m9ute or two ! fixation w be complete1 & ! film becomes ( a dirty br[n col\r4 ,x is !n left expos$ to ! air to get rid ( all traces ( osmic acid1 & may aft]w>ds be /a9$ z describ$ bel[4 #b4 ,treatm5t ) 99saturat$ aque\s solution ( corrosive sublimate99 7,muir's me?od74 ,! cov]-glass on :i* ! film has be5 spread1 is float$ be=e ! latt] has "t to dry1 film d[nw>ds on a saturat$ solution ( corrosive sublimate 9 a wat* glass = half an h\r4 ,! cov]-glass is plac$ 9 di/ill$ wat] & !n 9 alcohol to remove excess ( corrosive sublimate1 & !n /a9$4 ,a little c>e is requir$ :5 wa%+ ! film to prev5t x slid+ bodily (f ! cov]-glass4 #c4 ,by dry+ & pass+ rapidly ?r\gh ! flame ( a ,buns5 burn]1 exactly z 9 prep>+ specim5s ( sputum1 c4 7p4 #aaa74 ,?is me?od is h&y = ord9>y cl9ical purposes4 #d4 ,by keep+ ! cov]slips at a temp]ature ( ab\t #bjj ,f4 7,ehrli*'s me?od74 ,ehrli* uses = ?is purpose a /rip ( copp] ab\t two in*es wide & a foot long :i* is support$ on a retort /& 9 a horizontal position4 ,"o end is heat$ by a ,buns5's burn] b5ea?4 ,! po9t 9 ! copp] /rip at :i* ! temp]ature is at boil+ po9t is readily asc]ta9$ by dropp+ a little wat] on4 ,! po9t at :i* a drop ( wat] assumes ! sph]ical /ate indicates a temp]ature !re ( #bab ,f4 ,! cov]slips are plac$ an in* or two fur!r ?an ?is po9t1 & kept !re at a temp]ature ( ab\t #bjj ,f4 = "s h\rs4 ,,/a9+ ,,me?ods4 ,fre% blood may be /a9$ by mix+ ) ,f]ri]'s fu*s9e solution3-- ,fu*s9e #a grm4 ,di/ill$ wat] #aej c4c4 ,dissolve & add ,alcohol 7#hj p] c5t47 #ej c4c4 ,neutral glyc]9e #bjj c4c4 ,a spot ( ?is solution is mix$ ) ! blood on a slide by means ( a m\nt$ ne$le1 & cov]$ ) a clean cov]-glass4 ,! r$ corpuscles are slightly /a9$1 :ile ! nuclei ( ! :ite corpuscles are /a9$ a bright crimson1 & ! 8blood plates0 a deep p9k col\r4 ,/a9$ prep>ations may also be obta9$ by us+ 99,toison's fluid991 :i* s]ves also = dilut+ ! blood 9 ord] to det]m9e ! exact numb] ( r$ & :ite corpuscles pres5t by means ( ,g[]s' or ! ,?oma-,zeiss hmocytomet]4 ,x is prep>$ ?us3-- ,glyc]9e #cj c4c4 #a oz4 ,sodium sulphate #h grms4 #b drms4 ,sodium *loride #a grm4 #ae grs4 ,me?yl violet #be grm4 #d grs4 ,di/ill$ wat] #afj c4c4 #e oz4 ,x /a9s ! nuclei & blood plates1 b does n alt] ! %ape ( ! r$ cells4 ,x requires to be made up fre% occasionally z torul are apt to =m & multiply 9 x4 ,dri$ films may be /a9$ ) hmatoxyl9e1 picroc>m9e1 or any ( ! g5]al /a9s4 ,! nuclei ( ! leucocytes may be /a9$ rapidly 9 a c\ple ( m9utes 9 a "o p] c5t4 solution ( me?yl violet1 wa%+ 9 wat]1 dry+ betwe5 blott+ pap] & m\nt+ 9 balsam4 ,! be/ me?od = g5]al purposes is to /a9 ) a saturat$ aque\s solution ( me?yl blue = half an h\r or long]4 ,wa% 9 wat]1 & !n /a9 = t5 m9utes 9 a half saturat$ aque\s solution ( eos9e4 ,9 ?is way ! eos9ophile granules ( ! leucocytes & ! r$ corpuscles1 are /a9$ by ! eos9e1 :ile ! nuclei ( ! leucocytes are /a9$ by ! me?yl blue4 ,kan?ack & ,drysdale recomm5d t ! film %\ld fir/ be /a9$ ) a half p] c5t4 solution ( eos9e 9 #ej p] c5t4 alcohol1 !n wa%$1 dri$ & fix$ 9 ! flame1 & /a9$ = a %ort "t 9 ,lffl]'s solution ( me?yl5e blue 7p4 #ajd74 ,^! films may be /a9$ = micro-organisms 9 ! way describ$ = cov]-glass prep>ations 7p4 #aab74 ,,*apt] ,,viii4 ,,injection ,,( ,,blood ,,vessels4 ,injection ( blood vessels may be p]=m$ on small animals1 or on individual human organs aft] removal f ! body4 ,! object is to fill ! vessels ) a col\r$ fluid :i* w solidify aft]w>ds4 ,x is possible 9 ! same organ to inject ! art]ies ) a r$ m$ium1 ! ve9s blue1 & secretory ducts1 su* z bile ducts1 yell[ or blue4 ,! mo/ conv5i5t basis = an injection mass is gelat9e1 z its solutions liquefy at a temp]ature ( ab\t #ajj ,f41 & solidify a little bel[ t po9t1 & :5 solidifi$ cut readily1 & d n t5d to become brittle4 ,! v>i\s masses are prep>$ z foll[s3-- 99,r$ injection mass99 7,woodhead's =mula7 consi/s ( gelat9e s(t5$ by mixture ) wat] & col\r$ by c>m9e4 7#a7 ,c>m9e #d grms4 ,liq4 ammoni ,b4,p4 #h grms4 ,di/ill$ wat] #aej c4c4 ,dissolve ! c>m9e 9 ! ammonia 9 a mort>4 ,p\r on ! wat]4 ,mix ?or\ghly & filt]4 7#b7 ,gelat9e #aj grms4 ,di/ill$ wat] #ej c4c4 ,all[ x to /& 9 ! cold wat] until ! wat] is absorb$ & ! gelat9e has become s(t4 ,w>m 7#a7 almo/ to boil+ po9t ov] a ,buns5 burn]1 & add ! gelat9e sl[ly4 ,/ir ?or\ghly & add a t5 p] c5t4 solution ( acetic acid until ! solution becomes slightly acid4 ,?is w be %ewn by ! mass assum+ a d>k] & dull] col\r4 ,a little salicylic acid may be add$ to pres]ve x4 99,blue injection mass499--,to ! gelat9e mass 7#b7 prep>$ z above1 & liquefi$ by heat1 add in/ead ( c>m9e ,soluble ,prussian blue #e grms4 ,di/ill$ wat] #fj c4c4 ,e trace ( alkali mu/ be kept away f ! mass dur+ & aft] ! prep>ation4 ,sections ( inject$ organs %\ld be m\nt$ 9 ,f>rant's solution slightly acidulat$ ) =mic or acetic acid4 ,) e c>e1 h[ev]1 ! blue col\r is apt to fade 9 ! c\rse ( "t4 99,gre5 injection mass499 ,rob9's =mula 7modifi$74 7#a7 ,ars5iate ( soda 7sat4 sol47 #hj c4c4 ,glyc]9e #ej 7#b7 ,sulphate ( copp] 7sat4 sol47 #dj ,glyc]9e #ej ,mix & add "o "p to ?ree p>ts ( ! gelat9e mass made z = ! r$ & blue injections4 99,me?od ( injection499--,9 inject+ ! vessels ( tissues x is necess>y t ! organ or ! entire animal1 z ! case may be1 %all be kept dur+ injection at a temp]ature well above t at :i* ! gelat9e mass w melt1 o!rwise ! gelat9e w 8set0 9 ! art]ies & w nev] rea* ! capill>ies4 ,?is w>m+ is effect$ by imm]s+ ! animal 9 a wat] ba?4 ,! liquefi$ gelat9e is =c$ into ! art]y by a syr+e or by air pressure4 ,x is ess5tial t ! pressure be uni=m & /eady4 ,?is is s mu* m easily manag$ ) air pressure t ?is me?od is /rongly recomm5d$ to ! beg9n]4 ,b1 :atev] me?od be adopt$1 p]fect results c only be obta9$ ) c]ta9ty aft] long practice4 ,sometimes too high pressure w be employ$ & ! vessels give way1 at o!rs ! injection may n rea* ! capill>ies at all4 ,! mo/ scrupul\s att5tion to details is ess5tial4 ,by f> ! mo/ effective app>atus = inject+ is ! modification ( ,ludwig's con/ant pressure app>atus devis$ by ,fe>nley4#b ,al?\gh ! app>atus appe>s complicat$1 ! v>i\s p>ts are easily obta9$ & x w\ld be easy to improvise a sub/itute = ! wat] ba?4 #b 8,practical ,hi/ology10 7,macmillan ,co474 ,! app>atus :i* is %ewn 9 figures #aj & #aa consi/s ( a ba? deep en\gh to conta9 ! animal1 & a vessel conta9+ ! injection fluid4 ,! ba? is kept at a temp]ature ( ab\t #aaj by an ord9>y ,buns5 burn]4 ,a l>ge ,wolff's bottle 7#bj-dj oz47 ) ?ree necks1 is fitt$ ) ?ree india-rubb] /opp]s p]=at$ by glass tubes4 ,?r\gh ! c5tral /opp] a glass tube connect$ by a rubb] tube1 ) an ord9>y ,higg9son's syr+e1 passes almo/ to ! bottom ( ! bottle4 ,f "o ( ! o!r necks a rubb] tube passes to an ord9>y m]curial manomet]1 :ile f ! ?ird a tube passes to ! flask conta9+ ! liquefi$ injection mass1 :i* is imm]s$ 9 ! wat] ba?4 ,?is flask is also firmly /opp]$1 & %\ld be ab\t half fill$ ) injection mat]ial4 ,! deliv]y tube f ! l>ge ,wolff's bottle %\ld only j come ?r\gh ! cork4 ,ano!r glass tube passes d[n almo/ to ! bottom ( ! flask1 & is connect$ by a rubb] tube ) ! cannula ins]t$ into ! art]y4 ,x w be evid5t f figure #aa t :5 wat] is pump$ by ! ,higg9son's syr+e into ! ,wolff's bottle ! pressure !re w be rais$ 7z indicat$ by ! manomet]74 ,?is increase ( pressure w equally affect ! air inside ! bottle conta9+ ! injection fluid1 & ! fluid w be =c$ out along ! tube & ?r\gh ! cannula into ! art]y4 ,illu/ration3 ,fig4 #aj4--;,fe>nley's arrangem5t = inject+ blood vessels4 7,reproduc$ by p]mission ( ,messrs4 ,macmillan1 f ,fe>nley's ,practical ,hi/ology74 ,illu/ration3 ,fig4 #aa4--,s*eme %ew+ di/ribution ( pressure 9 ,fe>nley's ,injection ,app>atus 7f ,fe>nley's ,practical ,hi/ology74 ,be=e us+ ! app>atus a clamp %\ld be plac$ on ! exit tube ( ! vessel conta9+ ! injection fluid1 & ! pressure %\ld be rais$ to see t ! app>atus is ev]y:]e air-tight4 ,any leaks %\ld be seal$ be=e ! actual injection is comm5c$4 ,if an isolat$ organ is to be inject$1 a cannula ( glass or brass %\ld fir/ be ins]t$ into ! art]y & securely ti$ 9 position4 ,! organs1 if cold1 mu/ be soak$ 9 wat] at #abj ,f4 = ab\t half an h\r & !n transf]r$ to a wat] ba?4 ,9 ! case ( inject+ an entire animal1 su* z a rabbit1 rat1 or gu9ea pig1 ! injection is be/ made a few m9utes aft] dea?4 ,! animal may be *lor(orm$1 & !n bl$ to dea? by op5+ a l>ge ve94 ,z soon z dea? has occurr$ incise ! sk9 ov] ! ?orax 9 ! middle l9e4 ,cut ?r\gh ! co/al c>tilages to ! "r ( ! /]num1 & ?r\gh ! junction ( ! manubrium & body ( ! /]num4 ,^! incisions be+ = mo/ "p ?r\gh non-vascul> p>ts w n lead to escape ( fluid dur+ injection4 ,! /]num be+ =cibly rais$ t[>ds ! left1 ! p]ic>dium w be expos$ & mu/ be c>efully divid$4 ,an incision mu/ be made into ! left v5tricle1 & a cannula pass$ up into ! aorta & firmly secur$ by a ligature pass$ r\nd ! aorta ) ! assi/ance ( =ceps or an aneurism ne$le4 ,any blood is clean$ away & ! animal is !n plac$ 9 ! wat] ba? = ab\t t5 m9utes4 ,! tube f ! bottle conta9+ ! injection fluid is !n fill$ by g5tle pressure on ! syr+e1 & clamp$ :5 full4 ,its end is !n plac$ on ! cannula & secur$ !re by a ligature4 ,! pressure %\ld be rais$ by squeez+ ! syr+e until ! manomet] regi/]s "o in*4 ,! clamp %\ld !n be remov$ & ! injection comm5c$4 ,! pressure %\ld be rais$ v g5tly & con/antly by work+ ! syr+e1 & ! condition ( ! gums1 lips1 & eyes ( ! animal obs]v$4 ,! gums w soon %ew a p9k t+e4 ,! be/ indications are obta9$ by wat*+ ! effect on ! small vessels ( ! scl]otic4 ,:5 ^! are completely fill$1 :i* w be 9 ab\t five to t5 m9utes accord+ to ! rate at :i* ! air pressure has be5 increas$1 ! injection may be /opp$4 ,?is result w be obta9$1 "u good conditions1 be=e ! manomet] indicates a pressure ( five in*es4 ,! aorta %\ld n[ be ligatur$1 & ! animal plac$ 9 cold wat] frequ5tly r5ew$ until x is ?or\ghly cool$4 ,! organs may !n be remov$ & plac$ 9 me?ylat$ _s & h>d5$4 ,sections are aft]w>ds cut & m\nt$ 9 ! usual way4 ,,*apt] ,,ix4 ,,directions ,,= ,,prep>+ ,,individual ,,tissues4 99,normal hi/ology499--,x _c be too /rongly impress$ on ! beg9n] t a ?or\gh ma/]y ( ! normal appe>ances ( tissues & organs is absolutely necess>y be=e attempt+ to make an accurate /udy ( morbid *anges 9 !m4 ,he %\ld n be satisfi$ ) exam9+ "o specim5 ( an organ b z _m z he conv5i5tly c1 9 ord] to be fully acqua9t$ ) ! _m deviations f normal :i* may exi/ )\t actual disease4 ,he %\ld !re=e obta9 sev]al animals1 su* z small dogs1 cats1 rabbits1 frogs1 c41 & remove _! organs ) all c>e1 & h>d5 !m 9 ! v>i\s appropriate fluids4 ,he %\ld also obta9 specim5s ( normal human organs f ! po/-mortem room4 ,_m normal tissues 7sk91 muscle1 t5don1 bone1 c471 c also be prep>$ f a limb amputat$ = an accid5t to a heal?y pati5t4 ,by prep>+ specim5s 9 ?is way he w n only become ! possessor ( a set ( slides illu/rat+ normal hi/ology1 b w f9d also t he has acquir$ t pr(ici5cy 9 h>d5+ & /a9+ ! specim5s :i* practice alone c give4 ,! foll[+ acc\nt ( ! me?od ( prep>+ diff]5t tissues is m]ely int5d$ to indicate ! l9es on :i* ! beg9n] %\ld proce$4 ,aft] "s practice he w be q able to select ! modes ( h>d5+ & /a9+ :i* special circum/ances or cases may seem to dem&4 ,! fir/ "p ( ^! directions w ref] to ! prep>ation ( normal tissues1 ! second "p to morbid hi/ology4 99,blood499--,= special me?ods ( exam9ation see ,*apt] ,,vii4 99,blood cry/als--,hmoglob9 cry/als991 obta9$ f ! blood ( an animal1 or en\gh may be collect$ at any op]ation4 ,a little wat] or a little e!r is add$ to ! blood :i* is all[$ to /& = half-an-h\r aft] :i* a drop is all[$ to evaporate sl[ly on a clean slide4 99,hmat9 cry/als499--,! /ud5t %\ld make himself ?or\ghly famili> ) ^!1 z _! pres5ce af=ds positive pro( ( ! exi/5ce ( blood col\r+ matt] 9 a /a94 ,to obta9 !m a drop ( blood %\ld be all[$ to dry on a slide4 ,! dri$ blood is scrap$ into a little heap ) a small piece ( clean glass1 & a drop ( glacial acetic acid add$4 ,z x evaporates m9ute r$di%-br[n acicul> cry/als w appe>4 99,hmatoid9 cry/als499--,obta9$ f ! site ( a bruise1 or an old hmorrhage1 e4g41 a c]ebral apoplexy or a hmatocele4 99,simple squam\s epi!lium499--7,endo!lium74 ,c>efully /rip (f ! l9+ ( ! p>ietal p]ic>dium or p>ietal pleura1 ( a rec5tly kill$ animal1 or spread out its om5tum on a piece ( cork1 & 7#a7 /a9 ! int]cellul> cem5t ) nitrate ( silv] 7p4 #hb7 s z to reveal ! outl9es ( ! cells4 7#b7 ,/a9 o!r specim5s ) hmatoxyl9e or alum c>m9e to reveal ! nuclei4 99,/ratifi$ squam\s epi!lium499--,specim5s f sk9 ( v>i\s p>ts1 f+]1 gro91 lip1 tongue %\ld be prep>$4 ,h>d5 9 ,mll]'s fluid4 99,transitional epi!lium499--,occurs 9 ! pelvis ( ! kidney1 uret] & bladd]4 ,x is v readily deta*$1 especially if n h>d5$ imm$iately aft] dea?4 ,remove z e>ly z possible4 ,if ! bladd] is tak5 x %\ld be cut op5 & p9n$ out z flat z possible4 ,h>d5 9 osmic acid1 or ,mll]'s fluid & _s4 ,emb$ pref]ably 9 celloid94 99,simple column> epi!lium499--,occurs 9 _m p>ts4 ,x may be /udi$ 9 ! saliv>y ducts1 ! inte/9e1 kidney1 c41 ( any mammal4 99,goblet-cells499--,se5 abundantly among ! column> cells ( ! inte/9al gl&s1 & 9 ! muc\s gl&s ( ! m\? & ( ! c]vix ut]i4 99,/ratifi$ column> epi!lium499--,occurs only 9 ! ure?ra4 ,h>d5 ! p5is ( a cat 9 ,mll]'s fluid1 & cut transv]se sections4 99,ciliat$ epi!lium499--,h>d5 ! tra*ea ( a rec5tly kill$ cat 9 osmic acid or ,mll]'s fluid4 ,beautiful specim5s may also be obta9$ f an ord9>y nasal polypus1 :i* %\ld be put into h>d5+ fluid imm$iately aft] removal4 ,/a9 all sections ( epi!lium 9 picroc>m9e1 & 9 eos9e & hmatoxyl9e4 99,ord9>y areol> tissue499--,difficult to obta9 free f fat4 ,x may be /udi$ 9 ! subcutane\s tissue ( ! section ( ! cat's p5is already made4 ,a fragm5t ( ! tissue %\ld also be remov$ & c>efully teas$ 9 a drop ( picroc>m9e4 ,areol> tissue may also be /udi$ 9 sections ( sk91 & 9 ! capsules ( ! diff]5t int]nal organs4 99,ela/ic tissue499--,may also be /udi$ 9 mo/ sections ( sk94 ,if ! ligam5tum nu* ( a l>ge quadrup$ 7horse1 bullock71 c41 is available x yields ! be/ specim5s1 or ! human ligam5ta subflava may be exam9$4 ,p9 a piece out on a piece ( wood or wax4 ,h>d5 9 ,mll]'s fluid4 ,/a9 9 picroc>m9e4 ,bo? sections & teas$ specim5s %\ld be prep>$4 99,t5don499--,readily obta9$ f an amputat$ limb4 ,h>d5 9 ,mll]'s fluid4 ,make transv]se & longitud9al sections4 ,/a9 ) eos9e & hmatoxyl9e4 ,a prep>ation %\ld also be made by teas+ a little ( ! fre% t5don 9 normal salt solution1 & /a9+ ) picroc>m9e4 99,reti=m99 or 99lymphad5oid tissue994--,se5 9 lymphatic gl&s & 9 ! lymphoid follicles scatt]$ along ! sub-muc\s coat ( ! alim5t>y canal4 ,prep>e sections 9 ! ord9>y way4 ,/a9 9 eos9e & hmatoxyl9e or 9 picroc>m9e4 ,"s sections %\ld also be prep>$ by p5cill+ 7i4e41 dabb+ ) a camel's hair bru%7 or by %ak+ sections up 9 a te/ tube ) wat] or normal salt solution4 ,by ?is means ! leucocytes are remov$1 & ! /ructure ( ! ad5oid tissue itself becomes m evid5t4 99,fat499--,be/ /udi$ 9 sections ( sk9 & subcutane\s tissue1 or 9 ! mes5t]y ( ! cat4 ,"o specim5 %\ld be /a9$ ) osmic acid & picroc>m9e & m\nt$ 9 ,f>rant's m$ium1 & ano!r 9 eos9e & hmatoxyl9e & m\nt$ 9 ,canada balsam4 99,pigm5t cells499--,bran*$ cells are be/ /udi$ 9 ! liv+ foot ( ! frog1 :]e amboid movem5ts may be se5 9 !m :5 ! light fall+ on ! ret9a is made to v>y 9 int5sity4 ,p]man5t prep>ations are mo/ conv5i5tly made f ! pallium ( ! common snail4 ,! %ell is remov$1 & ! pallium snipp$ out ) ! scissors4 ,x is !n p9n$ out flat1 h>d5$ = a day 9 me?ylat$ _s1 & m\nt$ un/a9$ 9 ,f>rant's m$ium4 ,!y are also well se5 9 sections ( ! *oroid coat ( ! eye4 99,hyal9e c>tilage499--,specim5s may be obta9$ f any jo9t1 f ! co/al c>tilages ( "y animals1 or f ! ?yroid c>tilage & tra*eal r+s4 ,x may be h>d5$ 9 _s4 ,/a9 ) picroc>m9e1 eos9e & hmatoxyl9e1 & ) me?yl violet4 99,ela/ic c>tilage499--,prep>$ f ! epiglottis1 or f ! c>tilages ( ! e>1 e4g41 ( a cat4 ,h>d5 9 _s4 ,/a9 9 picroc>m9e or 9 dilute fu*s94 99,:ite fibro-c>tilage499--,obta9$ f int]v]tebral disc4 ,prep>e & /a9 z = hyal9e c>tilage4 99,bone993-- 99,uns(t5$ ,bone499--,cut z ?9 a section z possible ) a f9e saw4 ,rub ! section ) ! h& on a dry oil /one until x is z ?9 z possible4 ,!n cem5t x by ,canada balsam 7liquefi$ by w>m+7 to a piece ( plate glass & cont9ue ! rubb+ process ) ?is1 exam9+ x n[ & !n ) ! l[ p[] to see if x is ?9 en\gh4 ,z soon z x is ?9 en\gh x is wa%$ (f ! slide ) me?ylat$ _s1 & wa%$ to get rid ( ! f9e bone du/4 ,x %\ld !n be transf]r$ to turp5t9e & may be m\nt$ 9 balsam4 99,s(t5$ bone499--,specim5s may be obta9$ f an amputat$ limb or f ! femur ( a cat4 ,specim5s %\ld be decalcifi$ 9 *romic & nitric fluid1 & ! h>d5+ complet$ 9 _s4 ,9 /udy+ ! process ( ossification1 e4g41 9 ! head ( ! hum]us ( a kitt51 x is be/ to emb$ ! specim5 9 celloid9 be=e cutt+ sections1 z ! trabecul ( bone are v delicate1 & easily deta*$4 ,v beautiful d\ble /a9+ effects may be obta9$ ) ei!r picroc>m9e1 or eos9e & hmatoxyl9e1 & ) eos9e & me?yl violet4 99,bone m>r[499--,to obta9 good sections ( r$ bone m>r[1 take a piece ( ! clavicle or a rib1 or ( "o ( ! c>pal or t>sal bones4 ,decalcify 9 *romic & nitric fluid4 ,emb$ 9 celloid94 ,/a9 ) eos9e & logwood1 eos9e & alum c>m9e1 or alum c>m9e & picric acid4 ,m\nt 9 ,canada balsam4 ,! v>i\s cells pres5t 9 bone m>r[ may also be /udi$ by squeez+ "s fre% m>r[ f a rib1 & mak+ a cov]-glass film1 & prep>+ 9 exactly ! same way z is direct$ 9 ! case ( blood films on page #aaf4 99,too?499--,be/ cut 9 situ f ! jaw ( a cat4 ,decalcify 9 *romic & nitric fluid1 & cut bo? v]tical & transv]se sections4 ,/a9 9 picroc>m9e1 or eos9e & hmatoxyl9e4 99,develop+ too?499--,extremely good specim5s may be obta9$ f ! jaw ( a newly-born kitt5 or puppy4 ,sections c easily be made %ew+ a milk too? & a develop+ p]man5t too? by its side4 ,! enamel is dissolv$ by decalcify+ fluids4 ,to /udy x a specim5 ( uns(t5$ too? %\ld be made1 accord+ to ! directions giv5 = bone4 99,/rip$ muscle499--,%\ld be /udi$ 9 v>i\s animals4 ,! leg ( an insect su* z a cockroa* may be h>d5$ 9 osmic acid4 ,"o leg %\ld be h>d5$ 9 a /raight position s z to fix ! fibrils 9 ! fully ext5d$ position1 ano!r %\ld be b5t up s z to get specim5s ( relax$ fibrils4 ,portions ( muscle %\ld be remov$1 & teas$ on a glass slide 9 "s /a9+ fluid su* z picroc>m9e1 a t5? p] c5t4 solution ( eos9e or qu>t] p] c5t4 ( safran9e4 ,sections ( amphibian & mammalian muscle %\ld be prep>$ to %[ _! diff]5ces 9 /ructure4 ,! mo/ conv5i5t "p to select is ! tongue1 z a view ( ! fibres is obta9$ bo? 9 longitud9al & transv]se sections4 ,sections %\ld be /a9$ 9 eos9e & hmatoxyl9e :i* gives a beautiful effect4 ,= special /a9s = intra-muscul> n]ve end+s see page #ib4 99,he>t muscle499--,a portion %\ld be teas$ fre% 9 picroc>m9e or eos9e1 ano!r portion h>d5$ 9 ,mll]'s fluid1 & sections made & /a9$ ) eos9e & hmatoxyl9e4 99,un/rip$ muscle99 may be obta9$ by teas+ a fre% portion ( ! muscul> coat ( ! small inte/9e ( an animal1 or by sections ( ! h>d5$ inte/9e1 bladd] or ut]us4 ,/a9 9 picroc>m9e or pref]ably eos9e & hmatoxyl9e4 99,n]ves499--,! special me?ods = /a9+ n]ve tissues are detail$ 9 ,*apt] ,,vi4 ,! /ud5t mu/ rememb] t ! ord9>y /a9+ me?ods are also applicable to n]v\s tissues4 99,n]ve t]m9ations993-- 99,meissn]'s corpuscles499--,take ! tip ( an index f+] imm$iately aft] amputation4 ,place "p ( x at once 9 *loride ( gold solution1 & ! re/ 9 ,mll]'s fluid until x is h>d5$4 ,sections /a9$ ) *loride ( gold %\ld be m\nt$ 9 ,f>rant's m$ium4 ,! o!r sections may be /a9$ 9 picroc>m9e or eos9e & hmatoxyl9e4 99,pac9i's corpuscles499--,may be dissect$ out on ! small] bran*es ( ! digital n]ves1 or may be f\nd 9 ! mes5t]y ( ! cat4 ,! latt] %\ld be spread out on wood1 h>d5$ 9 ,mll]'s fluid1 /a9$ 9 hmatoxyl9e1 & m\nt$ 9 balsam4 ,o!r =ms ( tactile corpuscles may be /udi$ 9 ! tongues ( frogs1 ducks1 or geese4 ,a network ( n]v\s fibrils %\ld be /udi$ 9 ! cornea4 ,take ! cornea ( a newly kill$ frog or cat & /a9 ) *loride ( gold 7p4 #hb74 ,! end plates 9 :i* ! n]ves t]m9ate 9 muscle may be /udi$ by plac+ specim5s ( liv+ muscle ( "s cold blood$ animal into *loride ( gold solution1 & /a9+ r deeply4 99,art]ies499--,take a piece ( ! aorta1 a piece ( "s m$ium art]y1 z ! r5al or radial1 & h>d5 9 ,mll]'s fluid4 ,/a9 9 picroc>m9e & always 9 eos9e & hmatoxyl9e4 ,art]ioles are be/ /udi$ 9 sections ( ! v>i\s organs4 ,?us !y are se5 9 ea* ,malpighian body ( ! sple51 9 ! b\nd>y zone ( ! kidney1 & s on4 ,a longitud9al surface view c also be obta9$ by /a9+ & exam9+ ! pia mat]4 99,ve9s499--,remove1 h>d51 & /a9 9 ! same way4 99,capill>ies499--,may be v well se5 9 ! foot ( ! frog4 ,/un a frog by /rik+ its head1 or by *lor(orm+ x4 ,fix x on a piece ( c>d ) a ,v %ap$ not* at "o end4 ,tie "o ( ! h9d feet by means ( ?reads atta*$ to its toes s t ! web ( ! foot is g5tly /ret*$ ov] ! ,v4 ,! foot c !n be readily exam9$ "u a #a_/b in* objective4 ,! foot mu/ be bru%$ f "t to "t ) normal salt solution to keep x moi/4 ,! movem5t ( blood 9 ! capill>ies1 c41 c !n be /udi$ = an h\r or two4 ,aft] dea? ! mes5t]y %\ld be spread out on a piece ( wood1 & h>d5$ = a few days 9 ,mll]'s fluid4 ,/a9 ) eos9e & hmatoxyl9e4 99,lymphatics499--,comm5cem5t ( lymphatics 9 s]\s membrane4 ,/a9 a piece ( cat's om5tum 9 nitrate ( silv] 7p4 #hb7 = "s m9utes4 ,aft] wa%+ keep 9 glyc]9e = ab\t a week & !n /a9 9 hmatoxyl9e & m\nt 9 ,f>rant's m$ium4 99,lymphatic gl&s499--,! lymphatic gl&s ( ! neck ( ! cat may be us$4 ,h>d5 9 ,mll]'s fluid4 ,/a9 9 picroc>m9e1 eos9e & hmatoxyl9e4 99,sk9 & sweat gl&s499--,sections %\ld be made f pieces tak5 7a7 f ! sole1 7b7 f ! sk9 ( ! body1 7c7 f ! axilla ( an adult to /udy ! pigm5t4 ,h>d5 9 ,mll]'s fluid4 ,/a9 9 picroc>m9e or eos9e & hmatoxyl9e4 99,hairs & sebace\s gl&s499--,take a portion ( ! scalp1 or ( ! sk9 ( a puppy4 ,h>d5 9 ,mll]'s fluid4 ,/a9 9 eos9e & hmatoxyl9e1 & m\nt o!rs un/a9$4 ,hairs f v>i\s p>ts ( ! body %\ld also be soak$ = "s h\rs 9 liq4 potass & m\nt$ un/a9$ 9 ,f>rant's m$ium4 ,!y may be blea*$ subsequ5tly by treatm5t ) eau de ,javelle 7p4 #bg74 99,bra9 & sp9al cord499--,mu/ be remov$ f ! body ) extreme c>e1 all /ret*+ or squeez+ be+ avoid$4 ,h>d5 sl[ly 9 ,mll]'s fluid to :i* a f\r? ( its bulk ( wat] may be add$4 ,! be/ /a9+ reag5ts to employ are eos9e & hmatoxyl9e1 alum c>m9e or borax c>m9e1 anil9e blue-black1 c4 ,/a9+ me?ods1 see ,*apt] ,,vi4 99,eye499--,h>d5 ! eye ( a rec5tly kill$ bullock1 cat1 or o!r animal 9 =mal 7p4 #bc71 punctur+ ! scl]otic 9 places to all[ ! h>d5+ fluid to p5etrate4 ,9 ab\t a week make a horizontal section ?r\gh ! eye4 ,! ant]ior half 7! l5s hav+ be5 remov$7 may be satisfactorily cut 9 gum4 ,sections ( ! cry/all9e l5s are n v satisfactory4 ,! be/ way to get specim5s ( ! fibres is to tease a piece ( ! fre% l5s ( a fi% 7e4g41 a cod7 9 a #a_/dj p] c5t4 aque\s solution ( eos9e4 ,wa% ! eos9e (f ! slide ) #a_/b p] c5t4 acetic acid1 & m\nt 9 ,f>rant's solution4 ,! po/]ior half ( ! eye %\ld be emb$d$ 9 celloid91 z o!rwise x is extremely difficult to get sections ( ! ret9a 9 its prop] relation to ! o!r coats4 ,m\nt "s specim5s un/a9$4 ,/a9 o!rs ) ! ord9>y /a9s4 99,int]nal e>499--,decalcify+ ! temporal bone ( a cat1 dog1 gu9ea pig1 c41 9 *romic & nitric fluid4 ,z soon z ! bone is decalcifi$ complete ! h>d5+ ( ! s(t p>ts 9 me?ylat$ _s1 emb$ 9 celloid91 & cut sections 9 ! longitud9al axis ( ! co*lea4 ,ow+ to ! extreme h>dness ( ! bone 9 adults x w be f\nd be/ to use ! petr\s bone ( newly born animals4 ,! semi-circul> canals w be mo/ readily /udi$ 9 ! temporal bone ( fi%es1 or ( birds1 e4g41 ! common f[l4 ,!y also mu/ be cut 9 celloid91 & /a9$ 9 ! ord9>y way4 99,nose & olfactory epi!lium499--,x is difficult to obta9 specim5s f ! human subject1 b v satisfactory prep>ations may be made f ! dog1 or m conv5i5tly 9 a new born puppy :]e ! bones are /ill c>tilag9\s4 ,h>d5 ! latt] 9 ,mll]'s fluid1 decalcify adult specim5s 9 *romic & nitric fluid4 ,specim5s ( ciliat$ epi!lium1 c41 w be obta9$ f ! l[] "p1 & ( ! special olfactory epi!lium f ! upp] "p4 ,/a9 9 eos9e & hmatoxyl9e4 99,lungs499--,c>efully remove ! lungs ( a cat )\t injur+ ! bron*i or tra*ea4 ,introduce a cannula into ! tra*ea & g5tly inflate ! tra*ea ) air4 ,ligature ! tra*ea & place ! lung 9 ,mll]'s fluid1 a weight be+ atta*$ to keep ! organ subm]g$4 ,h>d5 = ab\t six weeks1 & !n make sections ( ! v>i\s p>ts4 ,to demon/rate ! endo!lium ( ! alveoli1 inject in/ead ( air1 nitrate ( silv]4 ,all[ x to rema9 9 = half an h\r1 !n remove x by wa%+1 & h>d5 9 ,mll]'s fluid4 ,beautiful ca/s ( ! alveoli1 c41 may be obta9$ by plac+ a cat's or human lung "u ! receiv] ( an air-pump1 & :5 ! air is completely exhau/$1 inject+ fusible metal into ! bron*us4 ,! lung tissue is !n remov$ by corrosion or by mac]ation4 ,portions ( ! ca/s %\ld be remov$1 fix$ 9 a glass cell ) a spot ( ,canada balsam1 & exam9$ by reflect$ light4 99,?yroid gl&499--,be/ obta9$ f a "y subject ei!r human or an animal4 ,h>d5 9 ,mll]'s fluid4 ,/a9 9 picroc>m9e or eos9e & hmatoxyl9e4 ,also /a9 sections 9 safran9e1 :i* /a9s ! colloid mat]ial1 & also picks out any colloid =mation 9 ! cells !mselves4 99,?ymus499--,remove f a ftus or a v "y animal1 & prep>e 9 ! usual way4 99,tongue499--,t ( ! cat or rabbit s]ves v well4 ,ord9>y transv]se sections %\ld be made1 & also sections ?r\gh ! circumvallate papill 9 ord] to /udy ! 8ta/e buds40 99,saliv>y gl&s499--,^? ( a cat or dog d v well4 ,sections %\ld be made f ea* ( ! ?ree gl&s4 99,/oma*499--,t ( ! cat or dog %\ld be /udi$4 ,! organ mu/ be remov$ imm$iately aft] dea? be=e any po/-mortem dige/ion ( ! coats has occurr$4 ,! /oma* %\ld be op5$1 wa%$ g5tly & p9n$ out flat1 ) z little /ret*+ z possible on a piece ( wood1 & h>d5$ 9 ,mll]'s fluid4 ,sections %\ld be made 7a7 longitud9ally ?r\gh ! c>diac end to %[ ! transition f ! sophageal to ! ga/ric muc\s membrane1 7b7 f a portion ( ! great] curvature1 7c7 longitud9ally ?r\gh ! pyloric valve4 ,eos9e & hmatoxyl9e =m ! be/ /a9 = ! alim5t>y canal4 99,inte/9e499--,prep>e 9 ! same way z ! /oma*4 ,make sections f 7a7 ! upp] "p ( ! duod5um to %[ ,brunn]'s gl&s1 7b7 ! ileum1 7c7 a ,pey]'s pat*1 7d7 ! v]mi=m app5dix1 7e7 ! colon4 99,liv]499--,make an injection ( "o specim5 ) c>m9e & gelat9 7p4 #abj74 ,h>d5 9 me?ylat$ _s4 ,o!rs %\ld be h>d5$ 9 ,mll]'s fluid & /a9$ 9 ! usual way4 99,kidney1 supra-r5al1 & pancreas499--,same prep>ation z = liv]4 99,sple5499--,h>d5 9 ,mll]'s fluid4 ,m\nt "o section un/a9$4 ,%ake ano!r up ) wat] 9 a te/ tube to %ew ! /ructure ( ! pulp4 ,/a9 o!rs 9 eos9e & hmatoxyl9e4 99,bladd]499--,mu/ be remov$ & p9n$ out imm$iately aft] dea?1 z o!rwise ! epi!lium w be mac]at$ (f4 ,consequ5tly x mu/ be tak5 f an animal1 z a cat4 ,h>d5 9 osmic acid4 ,cut 9 celloid9 z ! coats are v apt to become deta*$4 99,p5is99 & 99te/is994--,readily obta9$ f dog1 cat1 or rat4 ,/a9 ) eos9e & hmatoxyl9e4 99,ut]us1 ov>ies1 & ,fallopian tubes499--,may be obta9$ f ! po/-mortem room or f ! l[] animals4 ,h>d5 9 ,mll]'s fluid1 & make sections f ! c]vix1 ! body ( ! ut]us1 ! ,fallopian tube1 & ! ov>y4 ,/a9 ) eos9e & hmatoxyl9e4 99,embryological specim5s499--,= sy/ematic "w special manuals %\ld be consult$4 ,specim5s %\ld be h>d5$ 9 osmic acid or 9 ,mll]'s fluid1 & cut 9 celloid91 or p>aff94 99,cl\dy swell+499--,specim5s are obta9$ f organs ( subjects :o h di$ 9 ! e>ly /age ( "s fev]4 ,!y %\ld be always h>d5$ 9 ,mll]'s fluid1 z ! appe>ances alt] if ! tissue is kept 9 _s = any l5g? ( "t4 99,fatty deg5]ation499--,prep>e f pati5ts :o h di$ ( exhau/+ diseases1 phosphorus poison+1 c4 ,/a9 9 osmic acid4 ,m\nt 9 ,f>rant's m$ium & keep 9 ! d>k4 99,mucoid deg5]ation499--,/udy 9 goblet cells ( normal inte/9e or ( ov>ian cy/s4 ,!re are no satisfactory selective /a9s = muc94 99,colloid deg5]ation499--,occurs 9 ! ?yroid gl&1 9 ! tubules ( ! kidney 9 _m diseases1 & ! pro/ate ( ! old4 ,/a9 9 safran9e4 99,waxy or l>dace\s deg5]ation499--,be/ /udi$ 9 liv]1 sple51 or kidneys4 ,x %\ld be se>*$ = 9 p]sons :o h di$ f a long illness1 accompani$ by suppuration1 e4g41 ph?isis or bone disease4 ,m\nt "o section un/a9$1 /a9 ano!r 9 me?yl violet1 a ?ird 9 a weak solution ( iod9e1 & exam9e ! latt] at once bo? by transmitt$ & reflect$ light4 ,! iod9e /a9 is n p]man5t4 ,ano!r section %\ld be /a9$ 9 osmic acid1 foll[$ by me?yl violet1 z waxy & fatty deg5]ation frequ5tly co-exi/4 99,hyal9e deg5]ation499--,se5 9 ! art]ioles ( ! sple5 9 "s cases ( typhoid & diph!ria4 ,! ord9>y /a9+ me?ods mu/ be us$4 99,calc>e\s deg5]ation499--,occurs aft] fatty deg5]ation 9 gummata & 9 a!romat\s art]ies4 ,x also occurs 9 ! matrix ( ! co/al c>tilages aft] middle life4 ,m\nt "o section un/a9$ & exam9e if possible ) ! pol>iscope4 ,/a9 o!rs 9 safran9e4 99,pigm5t>y deg5]ation499--,may be /udi$ 9 br[n atrophy ( he>t1 nutmeg liv]1 c4 ,x is also se5 well 9 sp9al & c]ebral n]ve cells ( ! ag$4 ,h>d5 9 ,mll]'s fluid & m\nt sections un/a9$4 ,x w be unnecess>y to recapitulate ! me?ods = h>d5+ ! v>i\s diseas$ organs z ! directions = ! normal organs hold good4 ,if ! pres5ce ( micro-organisms be suspect$1 h>d5 9 me?ylat$ _s or absolute alcohol1 b z a rule bo? = diseas$ organs & tum\rs ,mll]'s fluid w be f\nd ! mo/ satisfactory reag5t = g5]al use4 ,x sometimes happ5s1 h[ev]1 t x is inconv5i5t to wait sev]al weeks1 until ! ,mll]'s fluid has h>d5$ ! specim5 suffici5tly1 be=e mak+ sections4 ,9 ?is case ! be/ plan is to make fre% sections1 or else to cut a slice ab\t "o-eigh? ( an in* ?ick1 & h>d5 = ab\t ?ree days 9 pl5ty ( me?ylat$ _s1 or 9 =mal 7p4 #bc74 99,tum\rs499--,mll]'s fluid %\ld be employ$1 unless a m rapid ag5t is requir$4 ,me?ylat$ _s may be us$ 9 ! case ( epi!lioma1 ad5oma1 c41 b = s>coma1 myxoma1 tum\rs conta9+ cy/s or mu* blood1 ,mll]'s fluid yields by f> ! be/ results4 ,,books ,,( ,,ref]5ce4 ,me?ods 9 ,microscopical ,anatomy--,:itman4 ,practical ,pa?ology--,woodhead4 ,textbook ( ,bact]iology--,crook%ank4 ,manual = ,physiological ,laboratory--,h>ris & ,p[]4 ,practical ,hi/ology--,fe>nley4 ,practical ,pa?ology & ,hi/ology--,gibbes4 ,j\rnal ( ,microscopical ,society4 ,me?ods & ,=mul--,squire4 ,! ,human ,bra9--,goodall4 ,practical ,bact]iology--,kan?ack & ,drysdale4 ,me?ods ( ,microscopical ,rese>*--,cole4 ,,index4 ,abbe's cond5s]1 #aa ,absolute alcohol1 #ba ,acetate ( copp]1 #hi ,air bubbles1 removal (1 #ef ,alum c>m9e1 #gf hmatoxyl9e1 #fh1 #gj ,amyloid deg5]ation1 #adi ,anil9e blue-black1 #id oil1 #ajb1 #ajh oil wat]1 #ajg ,app>atus requir$1 #a ,areol> tissue1 #acc ,bact]ia1 /a9s =1 #ajc ,balsam bottle1 #eh ,b>rett's logwood solution1 #fi ,bevan ,lewis's me?od1 #id ,bi*romate ( potassium1 #ag ,bism>ck br[n1 #ajd ,blea*+ solution1 #bg ,blood cry/als1 #acj ,blood1 me?ods ( exam9+1 #aac ,blood-vessels1 injection (1 #abj ,blue injection mass1 #aba ,bone m>r[1 #acf ,bone1 sections (1 #acf ,borax c>m9e1 #ge ,bra91 me?ods ( /a9+1 #id ,buckley's modification ( ,golgi's me?od1 #ii ,calc>e\s deg5]ation1 #adi ,canada balsam solution1 #fa ,c>bolic acid1 #bc ,c>m9e1 #gd injection mass1 #abj ,ca?c>t microtome1 #ci ,ca?c>t-,fraz] microtome1 #db ,c$> oil1 #h1 #fc ,celloid91 #cj ,cem5t+ cov]-glasses1 #fe ,*loral hmatoxyl9e1 #ib ,*loride ( gold1 #hb1 #id ,*romic & nitric decalcify+ fluid1 #bf ,ciliat$ epi!lium1 #acb ,circulation 9 frog's foot1 #ada ,cl>ify+ sections1 #fc ,cle>+ ag5ts1 #fc ,cl\dy swell+1 #adh ,clove oil1 #fc ,colloid deg5]ation1 #adi ,column> epi!lium1 #acb ,corrosive sublimate h>d5+1 #bc /a9+1 #ajj ,cov]-glasses1 cleans+ (1 #eg ,cov]-glass prep>ations1 #aaa ,decalcify+ solutions1 #bf ,dehydration1 #fc ,eau de ,javelle1 #bg ,ebn]'s solution1 #bg ,ehrli*-,biondi fluid1 #he ,ehrli*'s hmatoxyl9e1 #gj me?od ( fix+ blood-films1 #aaf me?od = tub]cle bacilli1 #aaj ,ehrli*-,gram me?od = /a9+ bact]ia1 #ajh ,ela/ic c>tilage1 #ace tissue1 #acc ,emb$d+ me?ods1 #bi ,endo!lium1 #aca ,eos9e1 #gb ,eos9e & hmatoxyl9e1 #gc ,epi!lial cem5t1 #aca ,e!r spray microtome1 #ci ,fces1 /a9+ = bacilli1 #aab ,f>rant's solution1 #ei ,fat1 removal f sections1 #ei /a9+ (1 #acd ,fatty deg5]ation1 #adh ,fe>nley's injection app>atus1 #abc ,f]ri]'s fu*s9e solution1 #aag ,flemm+'s solution1 #be ,flotation ( sections1 #ee ,fold$ sections1 treatm5t (1 #eh ,=mal1 #bc ,fre% sections1 #eb ,fu*s9e1 #ajd ,g5tian violet1 #ajd ,gibbes' /a9 = tub]cle bacilli1 #aaa ,gold *loride1 #hb1 #id ,golgi's silv] me?od1 #if sublimate me?od1 #ii ,gram's iodide solution1 #aje me?od = /a9+ bact]ia1 #ajg ,gre5 injection mass1 #abb ,gum1 #bi ,hmat9 cry/als1 #aca ,hmatoid91 #aca ,hmatoxyl9e1 ,ehrli*'s1 #gj ,kle95b]g's1 #gj ,s*u*>dt's1 #fh ,sihl]'s1 #ib ,weig]t's1 #hh ,hmoglob9 cry/als1 #acj ,h>d5+ processes1 #ae ,hyal9e c>tilage1 #ace deg5]ation1 #adi ,ice freez+ microtome1 #df ,imm]sion l5ses1 #h ,injection ( blood-vessels1 #abj pulmon>y alveoli1 #ade ,int]nal e>1 #adc ,inte/9es1 #adf ,iod9e solution1 #aje ,jung's e!r spray microtome1 #de ,kle95b]g's hmatoxyl9e1 #gj ,l>dace\s deg5]ation1 #adi ,li?io-picroc>m9e1 #gi ,li?ium c>m9e1 #gd ,liv]1 #adg ,lffl]'s me?yl blue1 #ajd ,logwood1 #fh ,lymphoid tissue1 #acd ,m>*i's fluid1 #bd ,m>r[1 #acf ,me?yl blue1 #aja1 #ajd violet1 #hc ,me?ylat$ _s1 #ai ,micro-organisms1 /a9s =1 #ajc ,microscope1 #f ,microtome1 ,beck]1 #di ,cambridge rock+1 #di ,ca?c>t1 #ci ,ca?c>t-,fraz]1 #db ,jung1 #de ,s*anze1 #dg ,swift's1 #di ,williams'1 #df ,m\ld = p>aff9 emb$d+1 #cf ,m\nt+ me?ods1 #ee ,mucoid deg5]ation1 #adh ,muscle1 #ach ,muir's me?od ( h>d5+ films1 #aaf ,mll]'s fluid1 #ag & =mal1 #bj & _s1 #bj ,neels5's /a9 = tub]cle bacilli1 #aaj ,n]ve cells1 /a9s =1 #id end+s1 #aci fibres1 /a9s =1 #hg ,nissl's anil9e me?od1 #aja ,nitrate ( silv]1 #hb ,nitric acid z h>d5+ ag5t1 #be decalcify+ ag5t1 #bf decol\ris+ ag5t1 #aaj ,normal salt solution1 #ec ,nose piece1 #i ,objectives1 #g ,oil ( b]gamot1 #fc c$>1 #h1 #fc cloves1 #fc origanum1 #fc ,osmic acid z h>d5+ ag5t1 #ba /a9+ reag5t1 #ha ,pal's me?od1 #hf solution1 #ij ,p>aff91 #cd ,picroc>m9e1 #gh ,pigm5t cells1 #acd ,pigm5t>y deg5]ation1 #aej ,plane iron microtome knife1 #db ,rapid h>d5+1 #aej ,ret9a1 #adc ,safran9e1 #he ,saliv>y gl&s1 #adf ,s*f]-,pal me?od1 #ia ,s*anze microtome1 #dg ,s*u*>dt's hmatoxyl9e1 #fh ,sihl]'s *loral hmatoxyl9e1 #ib ,silv] nitrate1 /a9 = n]ve cells1 #if epi!lial cem5t1 #aca ,sk91 #adb ,sp9al cord1 #hf1 #adb ,sple51 #adg ,sputum1 /a9+ (1 #aaa ,squam\s epi!lium1 #aca ,/a9+ me?ods1 #fg seq4 ,/oma*1 #adf ,/rip$ muscle1 #ach ,sulphuric acid1 #aje ,sweat gl&s1 #adb ,t5don1 #acc ,te/+ a microscope1 #ac ,?ymus gl&1 #ade ,?yroid gl&1 #ade ,toison's fluid1 #aah ,too?1 sections (1 #acg ,transitional epi!lium1 #acb ,tub]cle bacillus1 /a9s =1 #aaj ,tum\rs1 h>d5+ (1 #aej ,un/rip$ muscle1 #aci ,ur9e1 exam9ation = bacilli1 #aab ,ut]us1 #adh ,von ,ebn]'s decalcify+ solution1 #bg ,waxy deg5]ation1 #adi ,weig]t's hmatoxyl9e me?od1 #hh me?od = /a9+ bact]ia1 #ajf ,williams' ice freez+ microtome1 #df ,woodhead's injection mass1 #abj ,xylol1 #fa ,ziehl's c>bol-fu*s9e1 #aje