If you are working with frozen sections, AR is an unneccessary step.
If working with paraffin, you can lower the AR temp and keep it in for a longer time. Cover the container to avoid evaporation.
thanx for your reply.
I not using frozen sections. the mice are perfused with 4% Formaldehyde. spinal cord changed into 15% 24hr 30% sucrose 24hr made 60um sections using microtome.
kept in PBS with azide.
I cover the water bath with aluminum foil and before placing the tissue in citraconic anhydrate solution i checked the temp with normal mercury thermometer (95C) then I will insert the sections.
I will try to reduce the temperature and perform the same protocol.
I used 0.1M PB I had written it wrongly 1M in above post. please ignore that.