Counting B cells in hemacytometer - (Oct/12/2008 )
I am working on B-cell lines and I always count my cells before splitting them or before any treatment. I would like to ask you whether my way of doing so, is correct:
1.I take 10microliters of my suspension and add it on the hemacytometer
2.Count the four big squares
3.Take the average of these 4 big squares and
4.Multiply it by 10.000. The number I get is the number of cells/ml
So, I would like to ask you, if this procedure is correct, and why so I take 10 microliters and not another quantity??
Thank you very much in advance!!
The method is same as counting lymphocytes.
U might be taking 10ul because that is the approximate volume of the hemocytometer.
The number of B-cells will depend upon the volume of cells suspension U r taking (and the dilution of the cell suspension also if U r using Trypan blue also.)
Number of cells = average of 4 big-squares X dilution(if diluting) X volume of suspension
the suspension should fill the squares uniformly and should not flood the slide nor fall short.
It might be that U r taking 10ml of the suspension and for that reason multiplying by 10.
Number of cells = average of 4 big-squares X 10
Please see this thread and there is one PDF of cell-counting posted at 4th or 5th reply.
Thank you very much for yr immediate response!!!