Hello Constantin,

First step to your IHS protocol is fixing tissue on slide: dry slides overnight at RT, then rehydrate the following day and ensure the tissue is really adhered to the slides by bringing up to 95% ethanol, then immersing for 5 min in 95% ethanol / formalin (9:1 EtOH:37% formalin), then rinsing in 95% ethanol and hydrating down to water.

Second step: Continue to wash slides in warm water (37-40°C) will help you to remove gelatin (10-15 times).

Third step: Wash with PBT/Tween and block with PBT/TRITON/BSA/NGS.

(See also Cellular Pathology Technique C. F. A. Culling,R. T. Allison,W. T. Barr).

Hope it helps,

VM

Hello Constantin,

We work with Gelatin sections as well. Just keeping the slides at 37 C for 5 min (Needed you can keep it for upto 15mins) in PBS is sufficient to remove the Gelatin. 

At 37 C, Gelatin should dissolve completely in PBS and if the tissue fixed properly then I dont think, the tissue will get damaged at 37C 

Hi Constantin,

I have had the same problem as you, but this happened when my slides had already a year or so. As Shahul suggested I solved it having prolonged times (up to 1h) immersed in PBS at 37ºC . What also helped was to allow the slides to completely defrost in open air at RT (up to 2h) before putting in PBS.

Hope this helps,

Rita

Hi Constantin

PBS wash for three to five times (5 times) can remove gelatine from the coated slides for  frozen sections. 

But in case of Paraffin sections before washing with PBS you have to deparaffinize by using xylene, absolute alcohol, graded alcohol, distilled water and finally wash the slides with PBS. Through this procedures gelatine is thoroughly get washed and further you continue IHC procedures.

I would say you don't need to remove gelatin frpm slodes for immunocytochemistry.

 For what? If gelatin was pure and good quality - there will not be any unspecific stainings on slide. I was using such slides without any problems. Do you have unspecific stains??