Ultrahigh-resolution nonlinear fluorescence excitation microscopic system based on Bragg diffraction crystals

Abstract

The invention discloses an ultrahigh-resolution nonlinear fluorescence excitation microscopic system based on Bragg diffraction crystals, and relates to the field of laser detection. The system is combined with a nonlinear optical technology and a stimulated emission depletion microtechnique, infrared lasers with long wave lengths are adopted as a multiphoton excitation light source, a continuous laser beam is adopted as a depletion light source, the excitation light source is used for exciting fluorescent molecules, stimulated emission depletion light quenches fluorescence on the periphery of an excitation light spot focus, and the nonlinear optical microscopic system can break through the light wave diffraction limit. The Bragg diffraction crystals are introduced, the repetition frequency of femtosecond lasers is reduced below 4 MHz, and the fluorescence quantum yield of the fluorescent molecules is obviously improved. Meanwhile, the continuous stimulated emission depletion light is modulated into synchronization pulse lasers with the repetition frequency equal to that of the femtosecond lasers by the Bragg diffraction crystals, and on the basis of simplifying the device, the energy of incident light is reduced, and photobleaching on samples is reduced.

Description

technical field [0001] The invention relates to the field of laser detection, in particular to a super-high-resolution nonlinear excitation fluorescence microscope system based on Bragg diffraction crystals. Background technique [0002] The spatial resolution of conventional light microscopy has been limited by the diffraction limit. Due to its wave characteristics, light waves will be diffracted, so the beam cannot be infinitely focused, and 200nm is the theoretical resolution limit of conventional optical microscopes. With the rapid development of life sciences, the research has penetrated into the level of single cell, sub-cell and single molecule, and the spatial resolution of optical microscope has become the most critical core issue. [0003] In recent years, based on the development of modern measurement technology and the technological innovation brought by recent physicists, the far-field optical microscope has made revolutionary progress, and the resolution has b...

AI Technical Summary

Problems solved by technology

One of the bottlenecks encountered in the development of ultra-high resolution optical microscopy is that it is difficult to image living tissues

The essence is that these microscopic methods are based on single-photon excitation fluorescence technology, which has poor optical penetration and cannot meet various needs in practical applications.

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