Variable visual field scanning microscope and the method based on fixed light path system

Abstract

The invention discloses a variable-field scanning microscopic method based on a fixed optical path system, and a device thereof. At first, a beam expanding system is used for uniformly expanding laser beams which are transmitted from a laser so as to form uniform laser beams; then a two-dimensional space discrete system is used for the two-dimensional space discretization of the uniform laser beams so as to form a sub-beam of M multiplied by N; the sub-beam of M multiplied by N orderly passes through a collimating lens, a scanning galvanometer, a reflector, a dichroic mirror and a fluorescence microscope, and then focuses on the sample on a sample platform; thus the excited fluorescence can be formed on the sample; the fluorescence microscope is used for collecting the excited fluorescence, which is transmitted into an imaging system after being reflected by the dichroic mirror and focused by a focusing lens. The scanning of the galvanometer and the crossing movement of the sample platform are combined: the scanning step length and the scanning step number of the galvanometer are changed; the area of the unit viewing field is used as the step length to move the sample platform; thus the scanned viewing field can be changed. The scanning microscopic method realizes the flexible switching between the low-resolution large viewing field and the high-resolution small viewing field, and satisfies the requirements in different biomedical applications.

Description

technical field [0001] The invention relates to fluorescence microscopy technology, in particular to a scanning microscopy method and device for variable field of view based on a fixed optical path system. Background technique [0002] Fluorescence microscopy has become an important tool in life sciences, especially cell biology research. In recent years, with the development of life sciences, higher and higher requirements have been put forward for fluorescence microscopy technology. The continuous development of laser technology, fluorescent probe labeling technology, new fluorescence detection technology and imaging methods has become an important factor to promote the development of life sciences. important driving force. Laser scanning confocal microscopy and multiphoton excitation fluorescence microscopy can be used to achieve high spatial resolution three-dimensional imaging of biological samples. But their common disadvantage is that the sample needs to be scanned ...

AI Technical Summary

Problems solved by technology

In fact, limited by the nonlinear damage of the sample, the optical power required by the traditional single-point scanning multi-photon excitation fluorescence microscope to achieve sufficient photon number density is about 3-10mW

Therefore, the utilization rate of light energy is less than 5%

Due to the array point scanning, it is impossible for the MMM system to achieve electronic amplification in a single point scanning system, so its field of view and resolution are single, which limits its application in life science research

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