FCS Express: Proliferation Analysis

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Assays to assess the proliferative activity of cells are commonly used for monitoring the health and growth rate of cell populations.  In this type of assay, a cell tracking dye, such as CFSE, is incorporated into the cells.  With each round of cell division, the relative fluorescent intensity of the dye decreases by half.

As an initially homogeneous population of cells divides, you will get a mixture of  cells containing a half, a quarter, an eighth, a sixteenth, etc., of the initial population. Different conditions often cause a population of cells to change their proliferation characteristics, and understanding these effects is important in many different fields.

If you look at a histogram of this population after several divisions, you may see a plot like the one below:


Each peak (highlighted by an arrow) in the histogram above represents a population of cells that have divided. It is very common to have the different populations overlap, as they do above, because the standard deviation of the different populations is often greater than the separation between the populations. It is therefore impossible to simply choose a position and on the plot to denote a particular generation.

FCS Express uses a sophisticated linear least-squares fitting process to extract the correct information from your data and separate the populations.


As you can see, the fitting process extracted all of the different populations (green curves) and showed the resulting fit (red curve). In addition, you can see the data that was considered noise by the model (light blue curve).

Statistics


FCS Express provides a wide range of statistics about the results from the model.  An explanation of the different statistics can be found in our product manual.




It is possible to export any statistic from your proliferation fit to Excel, create PowerPoints slides, save layouts and more using our sophisticated . You can also use any proliferation statistic as a token in a text box or custom formula.

Formatting



FCS Express provides wealth of formatting options to control how your resulting histogram is displayed. The example below shows just some of the ways it is possible to format your results.


Model Options

FCS Express contains many options to allow you to control the parameters of your fit results.




FCS Express allows you to insert tokens into the Fixed Starting Generation and Background Noise fields. This means that you can effectively tell FCS Express to calculate the background from a control plot, and use that background setting for your proliferation analysis. As your data or gates change on the control  plot, your fit model will automatically update.


Try Proliferation Analysis for free for 30 days when you download a demo of FCS Express.