Hi all,
I am using animal-derived biological matrices as support for cell culture. At the end of the experiment I process the scaffolds+cells for histological examination. I include the samples either in paraffin or in a proper compound for cryopreservation. I then cut the samples to obtain sections about 8 micrometers thick. My problem is that the slices quite often detach from the slides during the staining procedure (I use glass slides, with positive charge, specific for histology).
Does anyone who works with these biomaterials have the same problem? Do you have any suggestion? I am looking for special treatments able to increase the stickiness of the slides.
Thanks in advance to anyone who will help!
Hi Elisa,
Have you tried poly lysine coating? or etching+silane treatment (this one may allow hydrophobic surfaces to attach your slides) of your slides?
Have you tried cryo sectioning, My colleagues at Uni work with decellularized nerve cultured with cells for histological sections. They are extra carefull when processing them,
Cryosectioning and embedeing them in OCT seems a better option as OCT evaporates after wards and you can stain it straigt away
i think decreasing the thickness to 5um helps. as well as using silane treated slides. another important thing in my experience is dehydration. multiple alcohol dehydration usually cause my sections to detach and if possible you can dehydrate in one step with acetone, that was helpful for me.
Thank you very much for all your suggestions. I have never tried silane-treated slides. I think I will try them!
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