Hello, I am now doing the HE staining of mouse skin paraffin section, but my slices always looks like not being well stained by eosin. I tried different staining time and use the dyes from different company, but the result always the same.
The skin was cut at 6µm,and before deparaffinization I put the slide in the oven at 56°C for 1 hour. This is my staining protocol: 2x7min xylene, 2x10min 100% ethanol, 5min 90% ethanol, 5min 70% ethanol, 5min ddwater. 10min hematoxylin, 2sec 0.1% HCl, 5min tap water, 3min eosin, 10sec tap water, 10sec ddwater, 10sec 70% ethanol.
The dermis seems different form others staining result, and I don't know why.
Please help me! Thanks!
The first one is my staining picture and the second is the brain HE staining using the same protocol. The last one is HE staining from the aticle "Protective Effect of Chitin Urocanate Nanofibers against Ultraviolet Radiation" .
Check PH of stains or manipulate in PH and see the difference
Or Try with other stain/ combination PAS/Alcian Blue, could be see the difference between your treatded and untreated samples.
I think your sections might be a bit too thick, which is why the epidermis and sweat gland structures look so dark. Try sectioning at 4 um, with a sharp blade.
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