FLIM for High Content Screening: Thinking Outside of the Box

摘要

The quantification of fluorescence lifetime has been used by spectroscopists for many years. More recently high throughput screening, which can be thought of as automated, batch spectroscopy, has increasingly used fluorescence lifetime based assays. High content screening's heritage is microscopy rather than spectroscopy - being based on quantification of images rather than bulk measurements of solutions. Microscopy has traditionally borrowed from spectroscopy as microscopy hardware evolves to allow measurements to be scaled from wells or cuvettes to the level of pixels in an image. A good example of this is the recent profusion of 'spectral-confocals' capable of performing spectral analysis at the level of a single pixel. Fluorescence lifetime analysis is another tool from the spectroscopist's toolkit that is starting to be adopted by microscopists in the guise of fluorescence lifetime imaging (FLIM). This aim of this article is to highlight some of the ways in which the use of FLIM is being explored in the biosciences and look forward to how FLIM may be used in high content screening as a hook for novel targets.