Method and device for strengthening total internal reflection fluorescence microscopic imaging by means of surface plasma

Abstract

The invention provides a method for strengthening total internal reflection fluorescence microscopic imaging by means of surface plasma of high-level secondary axial symmetry polarized beams. The method comprises the steps that spatial filtering is carried out on laser beams emitted by a laser through a pinhole filter, and the laser beams are straightened into parallel beams through a collimating lens; the parallel beams are incident into a polarization transition system to converted in the aspect of a polarization state; amplitude and phase modulation is further carried out on the obtained secondary axial symmetry polarized beams through a pupil filter and an annular diaphragm; the modulated secondary axial symmetry polarized beams are further reflected into a focusing objective lens with a high numerical aperture through a dichroic beam splitter and are incident to a three-layer structure of 'a glass substrate-a metal membrane-a sample'; excited fluorescence signals are reflected into the focusing objective lens through the three-layer structure, beam-expansion is carried out on the fluorescence signals by the focusing objective lens, the fluorescence signals are transmitted by the dichroic beam splitter, the fluorescence signals are focused onto a pinhole array plate through the focusing lens after filtered by a filtering plate, and light signals are converted into electric signals through a detector to be further processed.

Description

technical field [0001] The invention relates to surface plasmon-enhanced total internal reflection fluorescence microscopic imaging technology, in particular to a surface plasmon-enhanced total internal reflection fluorescent microscopic imaging method and device based on advanced sub-axissymmetric polarized light beams. Background technique [0002] Total Internal Reflection Fluorescence Microscopy (TIRFM) generates an evanescent field at the glass-sample interface by making the incident beam exceed the critical angle, thereby selectively exciting the fluorescent molecules near the interface and realizing the fine structure of cells. microscopic imaging. Total internal reflection fluorescence microscopy can not only help to further understand cell function, but also improve the signal-to-noise ratio of detection signals. However, when performing dynamic and rapid imaging of biological microstructures, the frame acquisition rate is relatively high, usually greater than 100 ...

AI Technical Summary

Problems solved by technology

However, when performing dynamic and rapid imaging of biological microstructures, the frame acquisition rate is relatively large, usually greater than 100 frames per second, and the fluorescence signal intensity needs to be further increased

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