Novel fluorescence lifetime microscopy imaging device and method based on optical additive heterodyne modulation

Abstract

The invention relates to a novel fluorescent lifetime microimaging device and method on the basis of optical addition heterodyne modulation. The device comprises a fluorescent exciting light path, a reference light path and a fluorescent imaging light path, wherein a cosinusoidal modulated high-frequency exciting light excites a fluorescent sample to generate the florescent light with the same frequency, and the florescent light images on a light-sensitive surface of an image intensifier; moreover, a reference light which has the modulation frequency which is similar to the exciting light frequency is also irradiated to the light-sensitive surface of the image intensifier; a fluorescent image and the reference light are superposed on the light-sensitive surface of the image intensifier to form a heterodyne image; and finally, a fluorescent lifetime image is synthesized through phase-locking amplifying and polar coordinate phase diagram analyzing. The fluorescent lifetime microimaging device and method have the advantages of high fluorescent lifetime measurement resolution, high imaging efficiency, strong antijamming capability and the like, and moreover, the technical requirements on a core part-the image intensifier are low.

Description

technical field [0001] The invention relates to a fluorescence lifetime microscopic imaging device, in particular to a device and a method for performing fluorescence lifetime microscopic imaging of biological and chemical samples by means of the optical addition heterodyne modulation principle. Background technique [0002] Due to the emergence of fluorescent protein molecules (FP), the use of fluorescent labeling and fluorescence measurement techniques to study the biophysical chemistry of living cells and biological tissues has become one of the hotspots in life science research. Due to the advantages of good signal selectivity, high sensitivity, and strong ability to resist background noise interference, fluorescence lifetime microscopy imaging technology has broad application prospects in the field of biomedicine. [0003] There are two main methods of fluorescence lifetime microscopy imaging technology. The first one is time-domain pulse method. The representative tech...

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Problems solved by technology

[0004] The present invention proposes a novel fluorescence lifetime microscopy imaging device and method based on optical addition heterodyne modulation, which inherits the advantages of high imaging efficiency and strong anti-interference ability of frequency domain fluorescence lifetime microscopy, and solves the problem of frequency domain fluorescence lifetime microscopy at the same time. Problem of low time measurement resolution in micro-imaging technology

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