I have a problem in IHC technique for normal breast tissue (control) specific in antigen retrieval step where tissue didn't remain in slide. This problem appears only in normal breast tissue slide .
Dear Shrouk, Use positively charged slides.Reduce the antingen retieval time or temperature. Generally the the problem should be the same for both the normal and pathological specimens.
Poly lysine coated slides may solve the problem. 2nd if you have only fat tissue in normal breast tissue you wont get anything after antigen retrieval, be sure to have some ductal part in normal breast.
I use postive slide of type citoglas . is it not good ? and antigene retrival step ( citrate buffer & tween20 at PH 6 for 30 mint at 100 c
If the samples detach from the slides, bake the slides with the section on a slide warmer at 62°C for 1h and let them air dry at room temp for 20 min before starting the IHC protocol. Paraffin will melt and strongly attach the specimen to the slide again. Also use Charged Adhesion Slides. Good luck!
since normal breast have large proportion of fat tissue due to which tissue section not attach firmly if they detaching then you should bake the slide at 50 degree celsius for 1 hours as addition steps in the same IHC protocol and then proceed forward.
- Badges
- Science topic
- Similar topics
- Medicine
- Oncology
- Cancer Research