Tissue ProcessingDr. Saket Kumar21st August 2012HISTOLOGY :It is te !ranc o" science #ic $ea%s #it te gross & microsco'ic stu$( o" norma% tissue .HISTOPATHOLOGY :It is te !ranc o" science #ic $ea%s #it te gross & microsco'ic stu$( o" tissue a)ecte$ !( $isease.Tissue "or stu$( can !e o!taine$ "rom: *io'siesAuto'siesHISTOT+,H-I./+S: The techniques for processing the tissues, whether biopsies, larger specimen removed at surgery, or tissues from autopsyso as to enable the pathologist to study them under the microscope.Protoco%s "o%%o#e$ in Histotecni0ues 1 1. 2ecei't & I$enti3cation2. La!e%ing o" te s'ecimen #it num!ering 4. 5i6ation7. De($ration8. ,%earing9. Im'regnation:. +m!e$$ing;. Section cuttinge$ in te surgica% 'ato%og( %a!orator( a>e a request form tat %ists te 'atient in"ormation an$ istor( a%ong #it a $escri'tion o" te site o" origin. Te s'ecimen are accessione$ !( gi>ing tem a number tat #i%% i$enti"( eac s'ecimen "or eac 'atient 5i6ation It is a 'rocess in #ic a s'ecimen is treate$ !( e6'osing it to a 36ati>e "or a 'articu%ar 'erio$ o" time in or$er to "aci%itate te succee$ing ste's.Te 'ur'ose o" 36ation is to 'reser>e tissues 'ermanent%( in as %i"e?%ike a state as 'ossi!%e. Te 36ati>e sou%$ !e 18 @ 20 times more in >o%ume ten te s'ecimen.Aims o" 5i6ation :1. It sou%$ 're>ent auto%(sis & 'utre"action o" te ce%%.2. It sou%$ 'enetrate e>en%( an$ ra'i$%(.4. It sou%$ ar$en te tissues 7. Increase te o'tica% $ensit(8. Sou%$ not cause srinkage or s#e%%ing o" te ce%%s9. =ust not react #it te rece'tor sites & tus must not intere"ere #it te staining 'roce$ure.:. It must !e cea' an$ easi%( a>ai%a!%e.Te !its sou%$ o" te siAe o" a''ro6imate%( 2 6 2 cm & 7? 9 micrometer in tickness "or o'timum 36ation to take '%ace.Tese !its are ten '%ace$ in meta% cassettes or ca'su%es #ic are ten '%ace$ in te 36ati>e.Tin( !io'sies or sma%% s'ecimen can !e #ra''e$ in a 3%ter 'a'er an$ ten 'ut in a cassette & 36e$.Sim'%e 5i6ati>es5orma%inTe most common%( use$ 36ati>e is 5orma%in .It is 're'are$ !( mi6ing 70 B 5orma%$e($e gas in 100 #C> o" $isti%%e$ #ater.Te resu%tant mi6ture is 100 B5orma%in.2outine%(D 10 B "orma%in is use$ #ic is 're'are$ !( mi6ing 10 m% o" 100 B "orma%in in ente$ !( a$$ing 11? 19 B metano%.4. 5ormation o" !%ack "orma%in 'igment D Aci$ "orma%$e($e ematin. Oter Sim'%e 5i6ati>esG%utara%$e($eOsmium Tetrao6i$ePottasium Dicromate=ercuric ,%ori$e Oter Sim'%e 5i6ati>es Fcont$.GPicric aci$ HenkerIs Jui$HenkerKs 5orma% FHe%%(Ks 5%ui$G*ouinKs 5%ui$,om'oun$ 5i6ati>es=icroanatomica% 36ati>es:Tese are use$ to 'reser>e te anatom( o" te tissue.,(to%ogica% 36ati>es:Tese are use$ to 36 intrace%%u%ar structures.Histocemica% 36ati>es :Tese are use$ to $emonstrate te cemica% constituents o" te ce%%.=icroanatomica% 5i6ati>es.10 B 5orma% sa%ine : It is a microanatomica% 36ati>e. I$ea% "or 36ation o" !rain..*u)ere$ "orma%in: Due to te 'resence o" !u)erD te 'H o" te so%ution remains at neutra% or near neutra%. As a resu%tD 5orma%in 'igment "ormation $oesnKt take '%ace. ,(to%ogica% 5i6ati>es -uc%ear 36ati>es : ,arno(Ks 5%ui$ ,%arkeKs 5%ui$ -e#comerKs 5%ui$ 5%emmingKs 5%ui$ ,(to'%asmic 5i6ati>es :,am'(Ks 5%ui$ 2egau$Ks 5%ui$Histocemica% 5i6ati>es:5orma% sa%ine ,o%$ acetoneA!so%ute a%coo%Deca%ci3cation: It is te 'rocess o" remo>a% o" te ca%cium sa%ts "rom te s'ecimen. Te >arious agents use$ "or $eca%ci"(ing are1 .-itric aci$.H($roc%oric aci$.5ormic aci$.Picric aci$.Acetic aci$.,itric aci$De($ration: It is te 'rocess in #ic te #ater content in te tissue to !e 'rocesse$is com'%ete%( re$uce$ !( 'assing te tissue troug increasing concentrations o" $e($rating agents.Te >arious $e($rating agents use$ are 1+t(% a%coo%AcetoneIso'ro'(% a%coo%Dio6ane

Te $uration o" te 'roce$ure can !e note$ $o#n as11. :0 B a%coo% @ 1 our2. :0 B a%coo% @ 1 our4. e.*enAene : It iscarcinogenic.M(%ene : It is most common%( use$. ,%oro"orm: To6ic an$ e6'ensi>e.

Im'regnation:In tis te tissue is ke't in a #a6 !at containing mo%ten 'araLn #a6 "or 9 @ ; ours .Te #a6 is in3%trate$ in te interices o" te tissue #ic increases te o'tica% $i)erentiation & ar$ens te tissue & e%'s in eas( sectioning o" te tissue.Te >arious #a6es #ic are use$ areD 1. ParaLn #a62. Para'%ast4. Para'%ast '%us7. Ge%atin8. ,e%%oi$in Nar containing mo%ten 'araLn #a6:+m!e$$ing :It is $one !( trans"ering te tissue #ic as !een c%eare$ o" te a%coo% to a mou%$ 3%%e$ #it mo%ten #a6 & is a%%o#e$ to coo% & so%i$i"(.A"ter so%i$i3cationD a #a6 !%ock is o!taine$ #ic is ten sectione$ to o!tain ri!!ons. T('es o" =ou%$s:A. LeuckartKs =ou%$s:L? sa'e$ !rass 'ieces #ic is '%ace$ in o''osing 'ositions & can !e mani'u%ate$ to increase or $ecrease te siAe o" te !%ock to !e 're'are$.*. G%ass or =eta% 'etri $ises :,. Oatc g%ass D. Pa'er !oats .LeuckartKs mou%$s :ParaLn !%ockSection ,utting :It is te 'roce$ure in #ic te !%ocks #ic a>e !een 're'are$ are cut or sectione$ an$ tin stri's o" >ar(ing tickness are 're'are$. Te instrument !( #ic tis is $one is ca%%e$ as a =icrotome. TYP+S O5 =I,2OTO=+S:.S%i$ing .2otar( .2ocking.5reeAing.*ase s%e$ge 2otar( =icrotome: It is te most common%( use$.A%so kno#n as=innotKs 2otar( microtome.In tis te *%ock o%$er mo>es u' an$ $o#n #i%e te kni"e remains 36e$.It is suita!%e "or cutting o" sma%% tissues & seria% sections can !e taken on it. Parts o" a =icrotome F 2otar( G :A. *%ock o%$er*. Kni"e c%am' scre#s ,. Kni"e c%am'sD. *%ock a$Pustment+. Tickness gauge5. Ang%e o" ti%t a$PustmentG. O'erating an$%e.Tissue Joatation !at: It is a termostatica%%( contro%%e$ #ater !at #it te insi$e co%oure$ !%ack. It is maintaine$ at a tem'erature maintaine$ 8 @ 9 $egree !e%o# te me%ting 'oint o" 'araLn #a6.+%ectronic tissue Joatation !at:Staining : Staining o" te section is $one to !ring out te 'articu%ar $etai%s in te tissue un$er stu$( . Te most common%( use$ stain in routine 'ractice is Haemato6(%in & eosin stain. Proce$ure :1. De'araLniAation #it 6(%ene.2. H($ration4. Oas un$er #ater7. Stain #it Haemato6(%in "or 18 min8. Oas #it #ater9. Di)erentiate #it 1 B aci$ a%coo%:. Oas #it #ater "or 10 min;. Stain #it 1B +osin "or 2 minesuse$ "or 36ing te sections on te s%i$es :A%!umin so%ution F =a(orKs egg a%!uminGStarc 'asteGe%atin=ountants : DPM F Distrene Di!ut(% 'ta%ate M(%ene G.,ana$a *a%sam ,o%o'onium resin Ter'ene resin Automation:Automate$ tissue 'rocessor: A%% te !e"ore mentione$ 'roce$ures u'to te im'regnation ste' can !e $one automatica%%( in a sing%eD unmanne$ instrument D #ic is te Automate$ Tissue 'rocessor.A$>antages : It 'ro>i$es constant agitation $uring e>er( ste' #ic ensures !etter 36ation & 'rocessing. It re$uces te #ork %oa$ & in turns im'ro>es te o>era%% out'ut o" te %a!orator(.Automatic stainer:Automatic stainer2e"erences:Te6t!ook o" =e$ica% La!orator( Tecno%og(. @ 2amnik Soo$.Histotecno%og( an$ ,(totecno%og(? =ani'a% Aca$em( o" Higer +$ucation.###.en.#iki'e$ia.orgtt':CCen.#iki'e$ia.orgC#ikiC5i%e:+m'(semaQHQan$Q+.P'gTank You R