we just want to do Flow cytometry in vivo, but the mouse epididymal adipose tissue was frozen at -80C, can we do it? and how to make cell fluids?
Hello Hu Huan,
If you didnt suspended your cells apropiatelly and preserved them by using DMSO, probably your will not be able to use the tissue for Flow. Usually you disgregate the tissue by using collagenase or other enzymes and then resuspend them in RPMI, or other apropiate culture media with 10% DMSO before frezzing or FCS at 10% DMSO . Hope this helps
Hi Hu Huan, what are you planing to see with FC? If specific antibodies for inflammation, or insulin signaling etc. it is a very bad idea to do FC from frozen sections... But if you are planning of lipid content you may give it a try..
For cell suspension, there are several protocols but it is not easy! If you aim for only adipocytes, you will need 150/200 um cell strainers.. If you aim stromal vascular fraction (immune cells, stem cells, etc.) then you will need another protocol and 70/100 um cell strainers.. So bear in mind it is not a very easy assay already and trying with frozen samples is not a good idea, sorry ;)
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