Simultaneous, non-interfering collection of optimal fluorescent and backscattered light signals on the MRC 500/600

Abstract

One of the most important attributes of the confocal LM is that it can be used for imaging living cells. Unfortunately, cells that contain fluorescent dyes are far less tolerant of the intense illumination characteristic of microscopical examination than unstained cells. As a result, it early became evident that every effort should be made to extract as much information as possible from every photon striking the specimen. This implies not only utilizing detectors with high quantum efficiency but also detecting the light which is back-scattered by (BSL), or passing through, the specimen in addition to any fluorescent signal. Both transmitted light and BSL carry information about the optical properties of the specimen and it is possible to detect this information without compromising fluorescent signal collection.