Flow cytometry in the past generally required highly trained individuals. However, the introduction of benchtop flow cytometers into clinical laboratories has resulted in the involvement of more junior staff while many hospitals rotate laboratory staff on a regular basis. Thus, the use of a properly controlled flow cytometer will provide additional confidence to individuals undergoing training. Instrument control procedures should involve daily calibration or be involved, at the very least, every time the instrument is switched on. Simple protocols are available to assist the monitoring of the laser, fluidics, and optics.
Daily calibration is usually performed using commercially available latex beads and biological controls that allow the operator to monitor: (1) light scatter and fluorescence peak channel coefficients of variation (CVs), (2) light and fluorescence peak channel drift, and (3) instrument sensitivity and the facilitation of compensation setup to adjust for spectral overlap. Furthermore, biological controls, particularly those that are "full process" controls, will assist both the training of staff and the monitoring of the staining process. For instrument control, several types of bead standard are available: (1) blank beads (type 0), (2) alignment (type I) beads, (3) reference beads (type II), (4) compensation beads (type II/III), and (5) calibration and antibody-binding beads (type III) (1).
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