Microscope Hardware and Software Delays Cause Photo-Toxicity

Abstract

Technological advancements in the areas of sample illumination, image acquisition, and image processing have significantly improved the speed and sensitivity of fluorescence microscopy. In particular, light emitting diodes (LEDs) coupled to transistor-transistor logic (TTL) circuits have reduced photo-bleaching and photo-toxicity by limiting sample illumination to the camera exposure time. Unfortunately, many microscopes still rely on bulb-based light sources that cannot be configured with TTL. Moreover, even when TTL is enabled in conventional microscope software, hardware and software delays can still contribute to photo-toxicity and lead to additional delays between subsequent images, introducing errors in time lapse image recordings. The goal of the present article is to highlight the significance of these issues.