1. incorporation of tritiated-thymidine. This thymidine
analogue is build in
the DNA that is newly formed. The higher the proliferative response to an
immunogen, the higher the number of counts in the respective culture will be.
2. by labeling cells with the carboxyfluorescein diacetate, succinimidyl ester (CFDAse) cell divisions can be evaluated making use of a flow cytometer. Each daughter cell will get half the fluorescence of the mother cell. That way not only the reaction to an immunogen, but also the originally reaction population can be calculated. It is even possible to track divisions after the cells have been put back to and retrieved from an an animal. And it is possible to use the dye together with surface markers to discriminate cell populations or combine it with intracellular (cytokine) staining.
3. comparable to the tritiated-thymidine, the non-radioactive thymidine analogue bromodeoxyuridine (BrdU) can be added to the the culture. This method, making use of a BrdU specific antibody, has now been made possible to measure the DNA synthesis together with surface markers and intracellular staining.
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