I use formalin fixed, paraffin embedded tissues of birds embryo to IHC-P using primary-Ab SSEA4 and secondary-Ab conjugated with HRP, but after incubation in citrate buffer in pressure cooker. I use gelatin coated slides, but after Ag-retrieval I usually find some of the sections been loosed.
I suggest, it seems reasonable to try polylysin-coated slides, as that's problem is pure adhesion...
Dear Belal,
Best thing is first to use Superfrost ++ object glass for coupes, second attaching coupes to superfrost ++ glas at 30-35 C followed by drying of the sections at least for 24 hours at 37 C.
Good Luck
Hi there, I have also had the same problem previously with brain tissue and found that using a microwave to heat up the citrate buffer and dipping the slides into the hot buffer worked just as well. It seemed to be ok as long as you keep the temperature up.
The best slide treatment I have found is Vectabond from Vector Labs. (http://www.vectorlabs.com/catalog.aspx?prodID=16) . You can prepare 500 slides from one bottle, which cost about £40, so it's much cheaper than Superfrost ++ and just as simple as preparing polylysine- or chrome gel-coated slides. I have used it for IHC following AR by both microwave or pressure cooking without any problems. In addition, the treatment works very well for sticking down thin (0.35micron) sections of resin-embedded tissue during various prolonged staining procedures and it performed much better then Superfrost ++.
Hi Belal, We have had the same problem with some wax sections of human tissues. Superfrost slides are a big help and also wrapping the slides carefully with a fine gauze reduced the problem. Also we use a microwave system rather than a pressure cooker so that might help.
I had different problems using microwave or cooker. I could not maintain desired temperature. Then I found a very easy and interesting technique. I warm the water in a beaker on hot plate, inside that beaker, we put 50 ml tube filled with citrate buffer. Wait till temperature in both reach up to 95 and then put the slide in the tube. Thus you save your sections from different temperature shocks and and also with this system you can continuously monitor the temperature and control it. Good luck
- Badges
- Science method