Fluorescence lifetime imaging method and device based on spectrum division technology

Abstract

The invention discloses a fluorescence lifetime imaging device based on a spectrum division technology. The fluorescence lifetime imaging device comprises a light source and a microscope objective forconverging exciting light emitted by the light source and exciting a sample to emit fluorescence, and the device is provided with a spectrum division module used for carrying out spectrum division onfluorescence collected by the microscope objective, a detection module used for receiving fluorescent molecules of different wave bands after spectrum division, and a counting module used for carrying out photon counting on the fluorescent molecules and calculating fluorescence lifetime information. Meanwhile, the invention further discloses a fluorescence lifetime imaging method based on the spectrum division technology. According to the invention, a spectrum division technology is introduced, and wide-band fluorescence signals are decomposed to different channels through the spectrum division module and are received by different detectors, so that the influence of dead time of a single detector on fluorescence lifetime measurement in a traditional method is greatly relieved, and the problem of photon accumulation effect of fluorescence lifetime measurement is relieved; and the effective detection efficiency of photons is improved, and the life imaging speed is increased.

Description

technical field [0001] The invention belongs to the field of fluorescence lifetime imaging, in particular to a method and device for fluorescence lifetime imaging based on spectral technology. Background technique [0002] In the field of biomedical imaging, super-resolution fluorescence imaging has always been the focus of biomedical research. Through special labeling of biomolecules, fluorescence optical microscopy plays an important role in observing subcellular structures. For fluorescence information, there are four basic physical dimensions, including intensity information, wavelength information (absorption spectrum and emission spectrum), lifetime information and polarization information. Due to the specificity of fluorescent labeling, high-contrast results of labeled structures can be obtained only by acquiring intensity information, so most current imaging techniques are aimed at intensity information. In addition to intensity information, wavelength information i...

AI Technical Summary

Problems solved by technology

However, this lifetime system based on confocal and TCSPC counters also has limitations, the most critical being the limitation of imaging speed

Due to the influence of the dead time of the detector and TCSPC, in order to avoid the impact of photon accumulation on the lifetime, the light intensity must be weakened, and at the same time, in order to obtain enough photon numbers to obtain lifetime information, the single-point detection time must be extended. Therefore, the imaging speed is limited, making it difficult to apply to real-time lifetime imaging

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