Hello,
I have soft-tissue samples from different organs stored at -80 (previously snap-frozen in liquid nitrogen). I would need to isolate cell nuclei and fix on slides and I need the nucleus structure to be preserved as much as possible for spatial genome analysis. Is there an easy way of mechanically disrupting the tissue and obtain a cell suspension without the use of enzymatic digestion?
I've found an old paper were tissue was gently "touched" on a glass slide several times but I have no idea if it could work. Any comments on this method?
Thanks!!!
There is a method used by Suzana Herculano-Houzel
and Roberto Lent for estimating numbers of neurons
of the brain.
http://www.jneurosci.org/content/25/10/2518
https://www.ncbi.nlm.nih.gov/pubmed/15758160
Regards,
Joachim
How about centrifugation techniques inoovated by De Duve C et al for cell subfractionning?
I strongly doubt that you are going io have intact nuclei -even more
I strongly doubt that you can obtain intact nuclei -even more spatially intact structures the way you have done it.
You could check this protocol, maybe it would help. I am using it for isolating nuclei from different tissues for single-nucleus RNA-seq.
http://www.nature.com/nprot/journal/v11/n3/full/nprot.2016.015.html
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