Hi,
Never thought about it however now I need to ask anyways :)
When I use 0.4% Trypan blue for counting of PBMC/lymphocytes (oh btw, does anybody still use Hayem solution today?) I do a 1:1 dilution with trypan:cells in PBS and then count them using Malassez chamber.
The question is should I count that 1:1 dilution when I calculating the final amount of cells in my sample? Should I multiply the final cell number X 2?
Thank you!
Hi Mark,
I am not familiar with a Malassez chamber. However, if you dilute your cells with trypan Blue and count them in a Hemocytometer, you multiply the final cell number by the dilution. In your case, 1:1 dilution means that you would multiply the final cell number by 2.
We multiply the final cell count by 2 as PBMC in PBS is diluted 1:1 with Trypan Blue (25 µL trypan Blue+25 µL PBMC in PBS).
I Have experience with Burker chamber, but I think the principle is the same for all chambers, so you have to take into account your dilution (and so multiply by 2)
Yes, whenever diluting you must count with the dilution factor, in your case 2.
Hi Mark,
I am sorry but I have never used Malassez chamber.
I usually use Neubauer chamber but I don't think it makes any difference.
You should always multiply your dilution factor, in your case 2.
Hope to be of any help
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