![]() ![]() ![]() When this enters the cell, it is cleaved to the active form CFSE, which is amine reactive, binding to intracellular proteins, and has been used extensively for cell tracking and proliferation. The most common of these dyes is carboxyfluorescein diacetate succinimidyl ester (or CFDA-SE). The first class is the intracellular dyes that enter the cell, are acted upon by cellular esterases which cleave the compound into a fluorescent form that can also interact with intracellular molecules, thus binding inside the cell. There are two major classes of these dyes, based on where the dye is retained by the cells. It segregates equally between daughter cells.The qualities of a good dye for proliferation include: Pick the right cellular proliferation dye.ĭetermine which dye you want to use for proliferation. To this end, there are several critical components in developing, validating and optimizing an assay to make these measures using flow cytometry. Proliferation of immune cells in response to stimulation.The focus here is on the long-term measure of cell division-the ‘temporal’ dimension for measuring such biological processes as: Proliferation calls for cells to make a trip around the cell cycle, and there are many ways to measure cell division. While these all can measure proliferation, they lack the finesse that flow cytometry can bring to the assay – which allows the phenotypic identification of which cells are actually dividing, as well as allowing for calculations of values such as the precursor frequency, the percentage of cells that have divided, a proliferation index, and more. More sophisticated assays can include the use of 3H-thymidine or colorimetric assays (MTT assays). This can be as simple as a cell count between untreated and treated cells. Measuring how cells proliferate in response to a stimulus is a time-honored assay in science. While normal cell proliferation keeps the body functioning, abnormal proliferation (such as in cancer) can be a target for therapy. Cellular proliferation is a critical component in biological systems. ![]()
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