Fsc-a Hot! Online

For a perfect sphere, FSC-A = FSC-H * FSC-W . If the cell is moving slowly, W increases, H decreases, but A remains constant.

If FSC-A is set incorrectly, your proliferation assays become noise, your cell cycle analysis becomes a lie, and your sorting purity plummets. This article dissects the physics, application, and troubleshooting of FSC-A to ensure your cytometric data is scientifically sound. To understand FSC-A, one must first understand what the "FSC" part means. Forward Scatter (FSC) detects light that passes through a cell and continues in a forward direction (typically 0.5° to 15° off the axis of the laser beam). Unlike Side Scatter (SSC), which detects refracted and reflected light at 90°, FSC intensity is directly proportional to the cell's surface area or diameter. For a perfect sphere, FSC-A = FSC-H * FSC-W

Use FSC-A for measuring the relative size of populations. Use FSC-H to check for signal saturation (if H maxes out, A may still be linear). Use FSC-W (in combination with A or H) for doublet discrimination . The Killer Application: Doublet Discrimination with FSC-A This is where FSC-A saves experiments. Flow cytometry assumes one event = one cell. However, two cells stuck together (a doublet) or three cells (a triplet) will pass through the laser and generate a single event. Unlike Side Scatter (SSC), which detects refracted and