The following images and text outline the basic steps involved in flow cytometric analysis as it occurs inside of the instrument.
1) Prepared cellular suspension enters the system via a sample probe.
2) The sample that is aspirated into the sample probe is dispensed into a flow cell.
3) Once entering the flow cell, the sample encounters isotonic sheath fluid that hydrodynamically focuses the cells so they exit the flow cell in single file / laminar flow.
4) As each individual cell exits the flow cell, it passes through a laser (or lasers).
5) While the cell is in contact with the laser, intrinsic and extrinsic characteristics are recorded via a series of dichroic mirrors and photo multiplier tubes. Forward scatter is determined by cell size, side scatter determines granularity, and the fluorescent signal reflects what MoAb was bound to the cell surface.
6) The cellular data is converted to an electronic signal, which can be sorted and analyzed in numerous ways via the system software.