Currently I am processing BrdU labelled rat brains for immunofluorescent detection. The cryosections are of 25 µm. My concern is that soon after the 2N HCl denaturation step the sections fall off the gelatin+chrome alum subbed slides.I tried with 0.5% gelatin upward till 3.0% and all have yielded similar result. Can anyone please help me out with this
Hi Arup;
Yes, as posted above thick sections fall easily, but I think you can use other options;
In substitution of your warm hand I will recommend you that use a hot plate at 37ºC , you will put the slide as son as you collect the specimen from the cryo in it for as long as 30 mins;
on the other hand you can use prepared slides with an adhesion treatment; in our lab use both options, the hot plate and this slides:
https://www.thermofisher.com/order/catalog/product/J1800AMNT?SID=srch-srp-J1800AMNT.
we use them for a variety of thickness and coronal/saigtal slices, with no problems at all.
You can try of course with " Aminoalkylsilane " an adhesive, you can find it here in ResearhGate,
Hope that help you!
Hi Arup, I recommand to use so called "Superfrost plus slides". You can use them for thick sections like 25µm and for stong tereatments like 2N HCl. We use them for our Insitu Hybridisation and they work pritty well.
Thank you all for your kind suggestions and help. We could troubleshoot the issue by using 2% APES coated slides. The section thickness was reduced to 20 micron
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