pisces22feb

Thanx for your reply. My sections are approximately 1cm and I do the xylene clearing untill the tissue is transparent and that too I give two washes of xylene.

hi, Can any one help me in deciphering the mistake in processing adult mouse skin and intestine for In Situ Hybridisation and histological staining. I am Fixing my tissue in 4% PFA overnight at 4 degree and then dehydrating in graded series of alcohol (25%,50%, 70%,80%,90% and 100%). Xylene is used as clearing for 3 minutes and 2min and then paraffin washes of 1 hour and overnight duration at 65degree, following embedding in paraffin. I am not getting good quality sections (5-7micro meters), tissue gets torn off from between the sections. Skin tissue section fall off from the slide during In Situ Hybridisation process.