wax-sectioning & staining-蚂蚁淘生物

FixsamplesinMEMFAandthenintoDent\"sFix(20%DMSO/80%Methanol)

Ifspecimensneedtobebleached,bleachinCurisBleachfor1/2hourunderwhitelight.Storespecimensat-20°Cin100%methanol

placeembryosinto100%ethanol(dehydratedwithmolecularsieve(type3A-BioRad)

infiltratespecimenswithparafin--placeembryosintoglasspetridishcontainingmoltenparaplast(58°-59°C).Repeatwithfreshmoltenwax.

fillabasemoldwith5mmparafin--moveembryosintomoldwithPipette

2Xwasheswithxylene,15minuteseach

Add50%EtOHand50%Hemo-De,20minutesforlargespecimens

Replacewith100%wax,2hoursat58°C

Replacewith100%wax,overnightat58°C*Alltimesareapproximate.Largesamplesrequirelongertime.

section

Heatsteelembeddingdishoverflamebrieflybeforeaddingwax(thispreventsbubblesfromformingandallowsevencoolingofthewax)

Placeenoughwaxtofillembeddingdish

Addspecimentodishandorientintodesiredposition

Placeembeddingringontheembeddingdish(makesurethatthereisenoughwaxinthedishsothattheembeddingringbecomesslightlyimmersedinthewax-thisholdstheringinplace)

Addhotwaxtothedishuntilitnearlyreachesthetopoftheembeddingring

Carefullyreorientthespecimenifithasmoved

Cooloncoolsurface,overnight

Removedishandsection

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