Imaging device and method for acquiring reflected signals and fluorescent signals simultaneously and imaging system

Abstract

The invention relates to an imaging device and method for acquiring reflected signals and fluorescent signals simultaneously and an imaging system. The device comprises an illumination light path, a beam splitting device, an objective lens and a detection light path, wherein the illumination light path is used for providing collimation and illumination beams; the beam splitting device is used for reflecting collimation and illumination beams with work wavelengths of reflected signals in a semi-transmissive and self-reflective manner and completely reflecting collimation and illumination beams with excitation wavelengths of the fluorescent signals; the objective lens is used for converging two light beams into a to-be-imaged surface of a sample, a reflection marker reflects the reflected signals, and a fluorescent marker is excited to reflect fluorescent emission signals; the reflected signals and the fluorescent emission signals reach the beam splitting device, and the reflected signals are transmitted in the semi-transmissive and self-reflective manner; the fluorescent emission signals are transmitted in a fully transmissive manner; the detection light path is used for detecting transmitted reflected signal and fluorescent emission signals respectively after separation. The reflected signals and the fluorescent signals are acquired simultaneously in the same system.

Description

technical field [0001] The invention relates to the acquisition of information of biological samples after staining, in particular to an imaging device, method and imaging system for simultaneously acquiring reflection signals and fluorescent signals. Background technique [0002] With the development of staining technology, more and more stains have been applied to the study of the structure and function of biological samples. Because different stains provide different cell structure or morphological information, multiple labeling methods are often applied to the study of the same sample structure and function, such as metal infiltration methods and fluorescent dye methods that can provide morphological or structural information, and can Gold nanoparticles, nanoshells, organic dyes, and quantum dots that provide molecular change information, among others. [0003] At present, a variety of optical imaging techniques have been developed to obtain reflection signals and fluor...

AI Technical Summary

Problems solved by technology

[0003] At present, a variety of optical imaging techniques have been developed to obtain reflection signals and fluorescence signals, but the current methods can only obtain reflection signals or fluorescence signals. For samples that need to obtain both reflection signals and fluorescence signals, most of them are through Different systems are used to detect separately, and they cannot be acquired at the same time, but can only be acquired separately. For example, Carson et al. invented a method to observe the fluorescence signal and the reflection signal in the same system. He switched the beam splitter ( 50 / 50) and a dichroic mirror to obtain a dual-mode (reflective and fluorescent) confocal microscope system (Carlson A L. et al. Journal of microscopy (2007), 228 (1): 11-24) , although this method realizes the observation of reflection signal and fluorescence signal in one system, it needs to acquire these two kinds of signals sequentially, which is not only inconvenient to operate, but also takes a long time to replace the beam splitter and two-way On the color mirror, and cannot automatically acquire the sample signal

[0004] To sum up, it is currently impossible to simultaneously acquire reflection signals and fluorescence signals of biological samples. Since both reflection signals and fluorescence signals can provide important sample structure and shape information, research on methods that can simultaneously acquire reflection signals and fluorescence signals technology is necessary

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