Fsc-a ~upd~ -
To get the best data, you need to set up your FSC-A correctly. The goal is to achieve optimal resolution without saturating the detector. Many modern cytometers use a parameter called . This is a factor that can be adjusted to ensure that the relationship between FSC-A and FSC-H is linear. The ideal result is a plot where the singlet population lies close to a 45-degree angle diagonal line. If the scaling is incorrect, you may see a "platform" effect where FSC-H values max out while FSC-A continues to increase. Adjusting the scaling factor helps correct this artifact. You should always check the voltage/gain for your FSC detector as well. Too low, and you'll lose the ability to distinguish small cells; too high, and your signals will be off-scale.
Doublets take longer to pass through the laser, disproportionately bloating the Area (FSC-A) relative to the Height (FSC-H), causing them to fall outside the diagonal line. To get the best data, you need to