Hello everyone,
After trypsin-EDTA treatment to the cells, I resuspend MDCK cells in 10ml of MEM (10%FBS). Then, once I load 10 micro-L of the suspension on Hemocytometer, I get clumps and very abnormal distribution in hemocytometer grids. I get them like 23, 12, 22 and 38, in 4 large grids of the hemocytometer. So, how possibly, I can improve my hemocytometer count?
Hi Mazhar Hussain,
To avoid clumps while harvest itself re-suspend the cells and post centifugation initially re-suspend in your cells in less quantity of media mix thoroughly and based on cell density (Visually) add more media. Even you can do more dilution before counting with hemocytometer like 1:100 etc.
For even distribution in hemocytometer - if your cover cells is moving after you charge your cells inside the chamber then you may likely get this type of distribution some times a small tissue paper debris too can cause this so try to avoid both you will definitely get a fine count
All the best
Regards
Sundarajan K
You can get some more valuable informations by following the links below:-
1. www.oglethorpe.edu/faculty/~k.../Lab1TissueCulture082808.doc
2. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2728112/
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