Cell nuclei count

Monitor progress of culture by taking 1-ml samples at least daily observe microscopically and carry out cell (nuclei) counts. 

If you want to see a cell nucleus, look down this microscope. On the spiritual side, if you want to see your Buddha nature, if you want to see Christ consciousness, if you want to see the religious side of the equation, fold your legs, sit down each day for two hours, count your breath from one to ten. Do that for five years and get back to me. ... 

Discard the supernatant and resuspend the cell pellet in 1 mL of HBSS/BSA. Mix 10 ul. of the cell suspension with 90 pi. of Kimura stain. With this stain, eosinophils have a green cytoplasm while mononuclear cells and neutrophils do not (all cells have a blue nucleus). Count the different cell types in a haemocytometer. Eosinophil purity should be >95% the predominant contaminating cells are mononuclear and the major exclusion criterion for the majority of our studies is the presence of >1% neutrophils. 

The interpretation of autoradiographs can be ambiguous, especially if the anatomy of the tissue furnishes few points of reference and the area to be counted is far from the cell nucleus. Often the shape of the cell may impede the exact determination of its centre . 

After injection, lyse cells and count the lysate by liquid scintillation. The volume injected into the nucleus of a mammalian cell is 1-2 X 10 ml, and up to 1-2 X 10 ° ml can be injected into the cytoplasm. A larger injection volume mostly kills the cells. 

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