An integrating averager of microspectrofluorometric signals

Optical signals are used to measure a variety of cell events; in particular, ratiometric fluorescent dyes offer diverse applications, but bring with them the difficult analytical problem of obtaining optical signals at rapidly alternating wavelengths. An improved method for using a computer to record these optical signals is described, and an example record of a Ca+2 transient given. A second technical problem is that ultraviolet (UV) light is required to excite many fluorescent dyes; many common lenses cannot transmit UV light. This article describes the tests needed to measure and optimize the optical path taken by light through a microscope and provides examples with sample lenses.