I've been using a standard H&E protocol without any issue on my cross-sections, however now I am using a swiss-roll protocol for my intestines and when I stain my slides, the tissue detaches from the slide. Does anyone have any insight as to why this may be occurring?
Many thanks.
Maybe you can smear your slide with white egg or albumin before you put the sample on the slide. It usually works
I usually heat my 7 um sections over night at 60 C..they look ugly coming out of the oven but once deparaffinized and stained they are perfect. I have used this technique for sections destined for IHC and IH applications with out issue.
Slides should b perfectly gelatin coated.dry slides smear at hotplate 60 c for15 min.then at room temperature overnight
Does your protocol include a period of incubation into the oven? For hystology, I generally use poly-l-lysine coated slides, which determine a stronger adhesion of your slices onto the glass. In addition, I usually put my fresh-cut slices into the oven at 37°C O/N or at 50°C for 1/2 hours before staining procedures.
Do you use coated slides, e.g. Superfrost? If not, you might apply a bit of albumin on your slides with your finger (as suggested by Siti Raudah) before you transfer your sections on the slides. Let them dry completely and after that staining shouldn't be a problem anymore.
Good luck!
You can check adhesive by finger touch whether sticky or not. Let the slide dry then deparafinize the slide in iven at 58-60°C for 2-5 min. Hope it will not detach.
If you choose to add albumin, it must contain thymol crystals (powdered) in order to avoid any fungal growth at later date. Also, addition of albumin may itself give false positive reaction especially after mucopolysaccharide staining, so try to avoid its use while demonstrating any proteins of mucopolysaccharides. If we take care of these points, albumin works. One more suggestion... try to avoid sudden and abrupt change of pH while shifting from one solution to next...adding few steps in between may consume time but will yield better results. Thank you.
http://www.ihcworld.com/_technical_tips/section_falloff_tips.htm
You must prepare your glass slides with Meyer albumine (from filtered egg and glycerine 1:1 and tymol crystals), and only after that you can lay your tissue sections.
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining