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Microtomy.pdf

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Microtomy.pdf

  1. 1. Date.3RaaAL xpt. No. PageNo.. *r*nssseara*****esrnsa" ICROTOMY REPARATIONDE fERMANENTSLIDE UE MaMMALIAN ISSUERAT INTODDCTON biecHo bestudie2undea micioscope nee prepouadim.f theohject issmallencughthroughahich ightcanpond,anðtha. Studyintinest detailk isnat uenpiseda whale moun prep.asuatimuil beSuficientln caae 0f laMge ojec they ae do_becukinthin Sectioastor the study his iscalled histolosica prepauatins ha objectiedhistological pHepauation is toundenstnnd th micro.anatomy 0t celli, TisSues,andongand and teCaralala Hha Structune dth suncdin Haematoayun A ~Asin Stoining pxctecol_is weameaant- tn histoloqyto examine Thin SectimA,Ahismehedknownas Double-Staining Method Teachers Signature..
  2. 2. Date No PagyeNo REQURED MATERLS Jfssues collecte Axom live specimen Bouin's uid (aqmeousonalcoholic- Distilled wcte ades of alcakoleg 5o,30%,q0, L100/) Xylal Para-tin :Haematoxylin and tosin Stains Kectanqulo block making maulds APX OH Canada balsam 7 ylam slides ove slips- Teachor's Sinnatu
  3. 3. Date pt No ROCEDUE A.CoLLECTION OEJissuE Fo Sectfoninq, issues are to be collectedfrom ive specimera. In Small vetebrates,the animal is paralysed by damaging te brair. Sn aqen vextebrates,the dame. Hesult isachieveo by staikinq heae aqainst a hrdobfec ha tssues anu cokécies immediate afte killing tho animal. FIXING Kfnq the tissue aithout anyStuuctuual ah chemical changa isknauan O&xing. Jhetissues ate Cut inte small pieces,lashed with phystological soluho eauiue, immersedinthe xativeandlost thea soradeiniliE boniodhemast (ommon fixative une3isBouin'sHuidauesuslalcehoi C ASHING Ohio is uDually doneaith Juate til t aeaqent ú empletely Hemaveà. SThepe1centage otwaten in the xative ia belous50,itghouldbe JAlashedonlyîn solutien Hecom.mended fothe puupose D DEHVDRATIONL he small pieces ot tissue aa cateied through dittxent guadesd alcahal,eg 5010,90,2 loo,HoDloLoen quae to ppe g9uade ond TM en ultimately toahsoulealcohol he_poxiod o tegbmentuih euchgade clependingondhe size oftissue luechangeAaa ivan inabsoluula olcahol. Teacher's Signature..
  4. 4. No E CLEARING Jhe tisu îs transfemed to Cecde. uoo oi, utMin 2 ous ha tiu ns thandpasentE and ana 1eady {or î'ntifratidn F INEILIRATION Jhis is to Heplace oil with paiatin. The intecelluluu Spare ane Completely occupied bypaattin he tissua. is ocahedinxylol andtranahehed to a taveed pn celain CYucible Containing Xulol and mo lten paraftin ar aHotit 1 and ha C3utbleis putinatempetature Contanalled poartin badh or o 1S minuteaAHerthopanio, thdissu fs transhe1ed to ancthen ave2 Caudecontaúning molttn tadfin anly, kep in hasome bath G. BLocK-MAKINGa PaaceDu2E Molten paxafin ispousredin_ablack_makingmould_andihetisauaixpu nth midale dt AHe dhaidening,extra poratin is treameðtn mave a Hectamgulan block_keeping the tis.sue in themiddle h SeeTioN CuTTIN hinis clone witha microiomeSectionson otine ulonKarauLally Ccutb-6Lmthick A queanefuee suide is tuken,oneauace is wmaxkedwitHaadiamona_pentl Adoplet ot Mayersalbumenissnat onthe mcnkedsduace. Athinfilm otdotenisput anthe slide andthu P.aa tin uibbon _aith sectionsaxaplaces on it.ke side isheated ona hot plala and the sechona ane propenly stmetched TM n h wate isdraned otf andthe ide isdried on the hot 8lal Teacher's Signature... ** * * * * * * * * * * * * * *
  5. 5. Date Page N 1 STANING OF HisToLOGICAL SLIOES ections of tissues ae usually Stained wi t tuao dye to oxdny CcntscasbE betuoeen dithetent histologca stsuuctudu.Ihis rna detciled study ecoie SAcining i tuoo dyes s knaen an doubla daining hi mos COnmon double SAaining practised in clam woHk 1s th heemaloryin and ensin. Jhe haematbryliw oeinqbasic dye împaul blua Caleuu t acidc mateyialb,i.e.,nucleicacids, whichaxo Cencentsucla in tha nucleuo andonly scaHeued 1nthe uoplaon lhe eocin isan acia due and the toplaomic mateials being bapic innatu a Stained hy it.Nho ceult a,he nucles andonu a smal 4uaciion Othe toplaom oppeav du, hil tha Heat d the ceall takesup s2colown. TM
  6. 6. Date No. Page No STAINING ECHNIQUE wha slde wit the panaffin aibbon Contodning Sections of . is dipped in xylol fon cabout 5 minuls. CAte the panatfin getcdissolvea,tho slide is transherdhed absoui alcehal Sh slide isthen Cannieo through qHades oaloho) from upper to lee. gMadeie joo,9o,t07A501 imeineach guadeor_altohal isabout 9-5 minuli. hen thaalide is transheroned to tasdistilledwate and net to tha haemataxylin Stain o_albout 30seconds to min At aDip_in. haemaiaxylinu,the uideiswashedln tapkat ) aenagaintha slidefscasthiedthraug gMadesofalcahal,hremlonert highe qrades orabaut 5-0minutes ineachgxade in Atenihat,the slide is then dipped in eosin fom too noua han theslide.is washedin90? aloholfollouedbytuoochangesinabzauli alcehol Afte Reeping the slidein Xylol foHabout minule,Hhe blideu Heady oH mounting kDASmal_amauntot Canada.balsom_o àPxisputan thaulide,depending onheSizececavenslip tobeuned JhiCouehslip s put oniE gi air bubbles ax lockedbetoeen thecoveneslipandthe slde,itmaybe Hemoved by leaving the slide oveHniGht ona hat plala. ail)hen the gide lsheady and istaatched unden compound mícrascap£e Hostudy thedetailed siudune ofhesue. Teacher's Signature.. 16
  7. 7. AvnaffinSection Mounhng on slide ylol Xylol lo aloho) 5min. qo% aluhol Casn) die loo/ altohol 0 min osin 90/o alcohol qo% odcoho) 10miw o alco hol 40,alcoho 1ominv S0 alcohei 50% alcohol tomin Distilled wa Distilled walu Alkaline wal Snin fou niny) Haematoxlin 3sec laP wal A A etJoaten Cuoas Citferentiatiar) FiG- Bouble staining, e hod (Suwmart)

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