Method: each detector/ camera is recorded at fixed settings using a range of sample intensities. Ideally, this is performed with a standardised external light source mounted at the objective, but could also be performed using the internal light sources and a mirror slide. Additionally, the Argolight check intensity pattern may be used. A minimum standard would be recording down to lowest detection sensitivity and up to saturation with at least three measurements between. Ideally, this would be performed across a range of detector gains, and using a calibrated external light source.
Justification: this allows monitoring of detector sensitivity and linearity, and with temporal sampling would allow early identification of faulty detectors (eg dying PMT, damaged camera vacuum chamber). These data, combined with excitation power at the objective, would allow accurate repetition of image acquisition parameters, improving data reproducibility within and between laboratories.
Requirements: external standardised light source or mirror slide. Argolight slide