Multi-fluorochrome flow cytometry experiments come with the drawback of “spectral overlap”. They have the potential to produce complex and extensive datasets, but only if this overlap is dealt with first.

Calculating and applying Compensation (to deal with the spectral overlap) is in theory simple, but a good understanding of the theory and application will ensure that the resulting data is correct.

During this module we look at the basic theory behind the overlap and compensation, how we calculate the transformation and how to ensure the matrix we arrive at is correct.

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