A system for measuring optical signal detector performance includes an optical signal detector comprising a first detection channel having a first light source and a first sensor. The first detection channel is configured to emit and focus light generated by the first light source at a first detection zone, and to receive and focus light on the first sensor. The system also includes a controller operatively coupled to the optical signal detector and configured to determine an operational performance status of the optical signal detector based on at least one of (i) a first measured characteristic of light focused on the sensor while a first non-fluorescent surface portion is in the first detection zone and (ii) a second measured characteristic of light focused on the sensor while a void is in the first detection zone. The optical signal detector can be a fluorometer.