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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 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).
StatisticsFCS Express provides a wide range of statistics about the results from the model. An explanation of the different statistics can be found in our manual. It is possible to export any statistic from your proliferation fit to Excel, create PowerPoints slides, save layouts and more using our sophisticated batch processing system. You can also use any proliferation statistic as a token in a text box or custom formula. FormattingFCS 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 OptionsFCS 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.
You can try Proliferation Analysis for free for 30 days. If you have never downloaded a demo before, you can use Proliferation Analysis in a free 30-day demo version from our web site. If you are an existing customer and wish to try the Proliferation module, please contact us and we will be glad to activate the proliferation module for you. |
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