I am facing an issue of debris after dissociation. These debris are mixed with dissociated cells. As I am using serum free and ultra low attachment flask, it is really hard to remove them. If you suggest to filter them out, what is the appropriate size of filter to use? Many site suggested to use 40 um. However, Eukaryotic cells are from 10 to 100 um in sizes. So, Is this fine with 40 um?
How to remove debris smaller than 40 um as those might still be mixed with cells? Thanks.
Hello
I have always remove cell debris by just allowing the cells to sediment in a centrifuge tube ....and then removing the supernatant. You will of course lose a small percentage of the cells. However centrifugation will only sediment the cells and debris
http://www.nature.com/nprot/journal/v3/n6/full/nprot.2008.65.html
http://www.sciencedirect.com/science/book/9780123854735
I have tried centrifugation before but it did not give satisfactory result. I tried it by different g forces e.g. 800g, 350g and 250g and neither one gave best result. Sometimes, tissue sample we received was so small e.g. 2.5 to 5 mm. So we also do not want to lose cells too.
Some debris were smaller than cells size and I am sure even we use 40 um filters, they will still be there.
Will it be possible to use ficoll paque to get rid of them? Anyone had tried before? Appreciate any suggestions if you have tried before and/or currently doing. Thanks.
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