Ultrasensitive flow cytometric analyses.

摘要

New techniques and approaches to cellular analysis being developed at the Los Alamos National Flow Cytometry Resource can be divided into those that improve sensitivity and those that move the technology into new areas by refining existing approaches. An example of the first category is a flow cytometric system capable of measuring the phase shift of fluorescence emitted by fluorophors bound to cells is being assembled. This phase sensitive cytometer is be capable of quantifying fluorescence life time on a cell-by-cell basis as well as using the phase sensitive detection to separate fluorescence emissions that overlap spectrally but have different lifetimes. A Fourier transform flow cytometer capable of measuring the fluorescence emission spectrum of individual labeled cells at rates approaching several hundred per second is also in the new technology category. The current implementation is capable of resolving the visible region of the spectrum into 8 bands. With this instrument, it is possible to resolve the contributions of fluorophors with overlapping emission spectra and to determine the emission spectra of dyes such as calcium concentration indicators that are sensitive to the physiological environment. Flow cytometric techniques have been refined to the point that it is possible to detect individual fluorescent molecules in solution as they flow past a laser beam. This capability has lead to a rapid DNA sequencing project. The goal of the project is to develop a technique that is capable of sequencing long strands of DNA (40,000 kb) at a rate of between 100 and 1,000 bases per second.