Compensation is an essential part of proper experimental setup for multicolor assays. However, it is also a subject that many flow cytometry users find difficult and confusing. Here we discuss some of the basics of spillover and compensation, tips …

Because the fluorophores used in flow cytometry emit photons of multiple energies and wavelengths, a mathematical method called compensation was developed to address the measurement of the photons of one fluorophore in multiple detectors.

3 Mistakes That Negatively Impact Flow Cytometry Experiments. Now that we’ve covered some proper methodology, let’s get to the 3 faulty methods that will bring confusion and inaccuracy to any flow cytometry experiment. Avoid the following practices at all costs. 1. Incorrectly employing a universal negative

For polychromatic flow cytometry, best practices in flow cytometry is to use the automated compensation methodologies. This will ensure consistent and accurate compensation, if some rules are followed.

Because the compensation matrix is more relevant to cytometry data — it tells us what to subtract from the detector in order to see the true fluorescence from its primary fluorophore — it is what is displayed in cytometry acquisition and analysis software.

However, through the use of appropriate single and double stained control samples, compensation for spectral overlap can be accomplished by electronic subtraction of unwanted signal, and accurate flow cytometric analysis of multi …

Figure 1.14.1 illustrates some of the com-mon errors in setting proper compensation. This unit is devoted to compensation in flow cytometry: why is it necessary, how is it accom-plished in hardware or in software, how com-pensation affects the visualization of data and, finally, how best to design an experiment to achieve proper compensation.

Jun 24, 2020 · In essence, AutoSpill uses iterations of linear regressions to get the best compensation or unmixing matrices, and removes the need for gating the positive and negative fractions of your controls. It saves you time, and generally provides better results. Learn more about AutoSpill here.

Data compensation is calculated from compensation matrix, which in turn, is calculated from spillover table. If the spillover table is stored in the form of a square matrix than its inversion generates compansation matrix.

Compensation has undergone a major redesign for FlowJo Version 10! Compensation in flow cytometry is the process of correcting for fluorescence spillover emissions. The detectors, or channels, in the instrument are designed to detect a very specific range of emissions.