I am working in a brain cortex region. I need to quantify the apoptosis during treatment. We don't have cryostat facility, So I need to follow the conventional paraffin embedded cortex sections. Is it possible to do tunnel assay with paraffin embedded tissues? If so please suggest me the suitable Tunnel assay kit (preferably fluoroscent).
Roche has a great kit for TUNEL staining in fluorescence and it works really well in FFPE tissues. You won't be disappointed.
https://www.sigmaaldrich.com/catalog/product/roche/12156792910?lang=en®ion=US
You can also get it from Fisher.
You may use this one < https://www.abcam.com/tunel-assay-kit-hrp-dab-ab206386.html >, its good
I have a lot of luck using TREVIGEN TACS 2 TdT - Blue Label Apoptosis Detection Kit for detecting tumor-induced apoptosis in mouse brain. I did some minor modification that turn it into fluorescent staining (see attached image).
https://trevigen.com/products-services/cell-stress-and-dna-damage/apoptosis/cell-stress-and-dna-damage-apoptosis-tunel-assay-kits/cell-stress-and-dna-damage-apoptosis-tunel-assay-kits-tacs-specialty-kits/tacs-kits/tacs-2-tdt-blue-label-kit/
The minor modification is done after step 11 of the attached protocol (page 11). Do not incubate in the Blue Label, instead, incubate in Alexa Fluor 488 Tyramide Reagent ( https://www.thermofisher.com/order/catalog/product/B40953) for 30 minutes, then DAPI for 5 minutes, then coverslip.
Hi Kamaladevi, I don't think so paraffin embedding is a good option particularly for brain tissues. The lipid content of the brain is unique from other tissues in the body because more than half of the brain contains membrane lipid. If you process for normal paraffin-based embedding method, you can lose majority of the lipid content in the brain tissue also morphology too. So its better to do cryosection after fixing with sucrose.
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