Recommended Literature Bastiaens, P. Fluorescence lifetime imaging microscopy: Spatial resolution of biochemical processes in the cell. Trends in Cell Biology 9: Levitt, J.
Triple Quadrupole LC-MS
HTRF technology | Cisbio
The application of time-resolved fluorescence spectroscopy TRFS to in vivo tissue diagnosis requires a method for fast acquisition of fluorescence decay profiles in multiple spectral bands. This study focusses on development of a clinically compatible fiber-optic based multispectral TRFS ms-TRFS system together with validation of its accuracy and precision for fluorescence lifetime measurements. It also presents the expansion of this technique into an imaging spectroscopy method. A tandem array of dichroic beamsplitters and filters was used to record TRFS decay profiles at four distinct spectral bands where biological tissue typically presents fluorescence emission maxima, namely, , , , and nm. A Laguerre-expansion deconvolution algorithm was used to compensate for modal dispersion inherent to large diameter optical fibers and the finite bandwidth of detectors and digitizers. Organic and biological chromophores with lifetimes that spanned a 0. Multi-spectral lifetime images of organic dye solutions contained in glass capillary tubes were recorded by raster scanning the single fiber probe in a 2D plane to validate the system as an imaging tool.
Though much of the interest in fluorescence in the past has been on measuring spectral qualities such as wavelength and intensity, there are two other highly useful intrinsic properties of fluorescence: lifetime or decay and anisotropy or polarization. Each has its own set of unique advantages, limitations, and challenges in detection when it comes to use in biological studies. This review will focus on the property of fluorescence lifetime, providing a brief background on instrumentation and theory, and examine the recent advancements and applications of measuring lifetime in the fields of high-throughput fluorescence lifetime imaging microscopy HT-FLIM and time-resolved flow cytometry TRFC. In addition, the crossover of these two methods and their outlooks will be discussed. The fluorescence lifetime is a unique optical parameter that has been exploited for many decades for a variety of biological applications.
Ewald Terpetschnig 1 and David M. Jameson 2 1 ISS Inc. University of Hawaii, HI The fluorescence lifetime is a measure of the time a fluorophore spends in the excited state before returning to the ground state by emitting a photon .