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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
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
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.
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
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).
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
You can also use any proliferation statistic as a token
in a text box or custom formula.
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.
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
proliferation analysis. As your data or gates change on the control
plot, your fit model will automatically update.