Lighting device and method

Abstract

The present invention relates to a lighting device and a method, particularly relating to a lighting device for improving axial resolution of a confocal microscope imaging system. The lighting device mainly comprises a collimating lens, an annular lens with a long focal length, an annular lens with a short focal length, and a converging lens whose one focal point is in a confocal imaging system lighting needle hole, wherein the focal points of the two annular lenses coincide. According to the device, the lighting light from the light source is shaped into annular parallel beams, after convergence, the beams illuminate a sample through the lighting needle hole of the confocal imaging system and an object lens. Compared with a traditional lighting mode, according to the device, the axial illumination to an object lens non-focal plane sample is reduced, the reflection light or emitted fluorescence from the non-focal plane sample collected by the confocal imaging system is reduced, and thus the noise of an image is reduced and the signal to noise ratio of the image is raised. Thus, the axial resolution of the confocal microscope imaging system can be raised by the lighting device.

Description

technical field [0001] The invention relates to an illuminating device and method, in particular to an illuminating device for improving the axial resolution of a confocal microscopic imaging system. The invention is mainly used in the field of biomedical microscopic imaging, and can also be used in material research and integrated circuit chip detection imaging. Background technique [0002] With the deepening of cell biology research, the application of fluorescence microscopy imaging is becoming more and more common, and confocal microscopy imaging has received more and more attention. The current confocal imaging system uses a traditional, uniform light source. The illumination light passes through the pinhole through the lens, and then illuminates the sample through the objective lens of the microscope. Finally, the detector collects the sample reflected or emitted by the objective lens and the pinhole. optical signal to obtain confocal images with a certain axial reso...

AI Technical Summary

Problems solved by technology

This shows that using conventional illumination methods, the illumination of the sample 17 located on the non-focal plane of the microscope objective lens 9 cannot be avoided, and the signal of the sample 17 located on the non-focal plane of the microscope objective lens is relative to the signal of the sample 10 located on the focal plane of the microscope objective lens. It is the image noise of the confocal imaging system, which reduces the signal-to-noise ratio of the image and reduces the axial resolution of the confocal imaging system

[0004] In addition, the confocal microscopy imaging system using a traditional collimated light source illuminates the sample 17 located on the non-focal plane of the microscope objective lens, which also speeds up the quenching of the fluorescent dye of the sample 17 located on the non-focal plane of the microscope objective lens

Therefore, when using a confocal microscopy imaging system for long-term illumination or three-dimensional scanning imaging of a sample, the requirements for anti-fade of fluorescence of the sample 10 located at the focal plane of the microscope objective lens and the sample 17 located at the non-focal plane of the microscope objective lens are higher, Increased the difficulty of scientific research experiments

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