Time-correlated single-photon counting using a confocal scanning laser microscope

摘要

Abstract: A system for 3D lifetime measurements of fluorophores has been developed and applied to multiple stained specimens. The system is built around a picosecond detector system for time correlated single photon counting and a confocal scanning laser microscope. It is capable of measuring lifetimes in the interval 1 - 20 ns, with a resolution of approximately 10 ps. By studying the lifetime of the fluorescence in a specimen it is possible to identify different fluorophores even if their emission spectra overlap. In addition, information regarding the chemical environment may be obtained due to effects from the environment on the fluorescence lifetime of the fluorophore.!5