Hello,
I need guidance in counting lymphocytes such as B cells or T cells using a light microscope.
What magnification should I be using? How big are the cells if I wanted to count them using a hemocytometer? Is there a good stain besides Trypan Blue to help count these types of cells and to determine viability? A link to a protocol for this procedure would be infinitely useful.
Well. Abdullah. You would love to know more about the morphology of the cells. Sizes. Stained of the cell. The shape and so on. Thus. It would be easier for you to count the cell and identification. Don't worry. Keep studying. Ask your lecturers.
Hi Abdullah
Your question is not very clear. Do you want to count the total lymphocyte population in the specimen or only the living cells? If you want to do the first thing, I suggest you make a smear, fix it and stain it with May-Grunwald-Giemsa stain. Lymphocytes are mononuclear cells with round nucleus and variable amount of cytoplasms. The nucleus of the small lymphocyte is around 7 microns (same size as a red blood cell).Most of them have scanty cytoplasm. If on the other hand you want to know how many of them are alive or dead, then you can use trypan blue but it would be difficult to get clear cytoplasmic details though. If you need any more help, I suggest you refer 'Practical Haematology' by Dacie and Lewis. Hope this helps you. Good luck. Siva
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