- Paraffin embedded tissues sections should be cut at about 7 micrometer thicknesses and placed on uncoated glass microscope slides with a drop of water to flatten. The sections are then baked at 60 C overnight to aid adherence of tissue to slide.
- Baked sections are deparaffinised in two 5 minute washes in xylene with agitation.
- Following deparaffinization, sections are rehydrated through ethanol series (2X100%, 95%, 70%, dH2O) for 2 minutes in each wash.
- Sections are then stained for 6 minutes in Haematoxylin and subsequently washed for 2 minutes in running H2O.
- Stain is differentiated for 30 seconds in acid alcohol, rinsed for 1 minute in running H2O and then further differentiated in ammonia water for 10 seconds and finally rinsed in H2O for 10 minutes.
- Sections are then stained for 6 minutes in eosin and dehydrated through 95% ethanol ( a few dips), 100% ethanol (3 X 2 minutes with agitation) and then cleared in two 2 minute washes of xylene.
- Slides should be coverslipped and sealed with Cytoseal (Fisher) for observation.
The Pleurogene Project
