生物實驗技術：Mollusk Histopathology Procedures

MolluskHistopathologyProcedures

1.Thereare30oysterinastandardsample.Theyarescrubbedfreeoffoulingorganisms,separatedfromoneanother,andlayedoutonatray.

2.Eachoysterismeasuredfromthehingetothebillinmillimeters,followingthecurveoftheshell.

3.Oystersareshuckedfromthehingetoavoiddamagingdelicatetissues.

4.TheoysterrectumisexcisedtocultureforPerkinusmarinus,whichcausesdermo-disease.

5.TubesofRay’sfluidthioglycollatemediumwithincubatingrecta.

6.Acrosssectioncontainingmantle,gills,digestivesystem,andgonadiscutfromthesamelocationineachoyster.

7.Crosssectionsarefixed,dehydrated,infiltratedwithmoltenparaffin,andembeddedinblocksofparaffin.

8.Eachtissuecrosssectionismovedfromitslabeledcassetteintoamoldfilledwithmoltenparaffin.

9.Thelabeledcassetteisplacedatopthemoldanditisplacedonachillingplatetosolidifytheparaffinblock

10.Ribbonsof5-6micronthicksectionsofthecrosssectionarecutfromtheparaffinblockonaspecializedinstrumentcalledamicrotome.

11.Theribbonisfloatedonawaterbathtospreadthesectionsandindividualsectionsareattachedtoglassslides.

12.Unstainedparaffinsectionsofrazorclamtissuesdrying.

13.Paraffinisremovedfromtheslideswithsolvents.ThetissuesareroutinelystainedwithMayer’shematoxylin(purple)andeosin(orange).

14.Apermanentprotectiveglasscoverslipismountedoverthestainedsection.

15.Finished