Hi,
I have isolated adipocytes from adipose tissue and intend on collecting conditioned medium from them. However, I need to standardise the concentration of the proteins in the conditioned medium by counting the adipocytes which is difficult because they float. Is there a protocol for using a haemocytometer to count adipocytes?
Thanks
Hi Melissa
I am not sure I understand your point.
To count cells, you have to individualise cells first if necessary (make a single cell suspension using trypsin, accutase or ...), then resuspend your cells in a known volume of medium or PBS, take an aliquot and count. You will end up with a number of cells per ml (after making the appropriate calculation depending on the type of your counting chamber). Knowing how much volume you have in total and considering the number of cells per ml, you can have your total cell number.
In your case, if your cells are already floating and as single cells, it is even easier. I would collect the medium and the adipocytes in a centrifuge tube, adjust the volume to a fixed amount of medium. Then Take an aliquot (add Trypan blue if you want to exclude dead cells) and count. You may need to perform a dilution or concentrate a bit your suspension.
please let me know if something is not clear or if I missunderstood your question.
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