Compensation is the process of correcting for spillover when one fluorophore is detected in multiple channels. It is required for most experiments of four or more colors to identify the correct signal that should be measured in each channel. This quick list can help...
When designing a flow cytometry experiment, it is important to consider the relative brightness of each fluorescent label on your specific instrument. Generally, it is best to assign brighter fluorochromes to weakly expressed markers, and dimmer labels to strongly...
Flow cytometry can be performed directly using a fluorescently labeled primary antibody, or indirectly using an unconjugated primary antibody with a labeled secondary. Most researchers prefer direct detection due to its simplicity and ease of use. However, certain...
The inability to resolve signals with overlapping color spectra has traditionally limited fluorescence microscopy to a “color-barrier” of 5 colors. However innovative new developments in microscopy techniques and analysis software are allowing researchers to resolve...
Flow cytometry cell cycle analysis typically involves using a DNA binding dye to determine each cell’s total DNA content. Sorting cells by cell cycle phase is then possible, as the total DNA content fluctuates as cells pass through G0/G1, S, and G2/M phases. Cells in...
The first step to any successful flow cytometry data analysis is proper gating. For a complete overview of gating terminology, types and strategies, check out our previous Gating Newsletter. Here we outline 6 questions that cytometry researchers should ask themselves...
