how can we detect the rate of B-cell replication?

Question

the method by which the rate of cell cycle can be detected.

Answer 1

The easiest method is to synchronize the cells and then label them with a quantitative membrane-binding dye (such as CFSE), the intensity of which would be reduced by two with each cell division (the daughter cells will inherit half of the mother cell's membrane each). Then you analyse cells by flow cytometry in time course and see when most cells halve their dye content.

You can also use a DNA-binding dye (such as PI) and measure the DNA content of your cells in time course. The DNA content will increase over time and then drop back to a minimum, which would mean that the cells entered the next cell cycle. The disadvantage of this method is, however, that you have to take probes often enough as the second and subsequent cell cycles will be indistinguishable - and you can in theory miss the second one.

Measuring cell cycle rates in an unsynchronised population is done technically in a similar way, but will require some mathematics, a so-called precursor-product relationship analysis. Using this method and radiolabeled glucose, it has even been possible to measure the cell cycle rate of T cells in humans (as described in the paper cited below)!

Answer 2

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