Real-time three-dimensional laser fluorescence microscopic imaging device

Abstract

The invention relates to a real-time three-dimensional laser fluorescence microscopic imaging device. The real-time three-dimensional laser fluorescence microscopic imaging device is compatible with a traditional bright field illumination microscopic imaging working mode. The real-time three-dimensional laser fluorescence microscopic imaging device is divided into a laser excitation module, a bright field imaging illumination module, a digital detection module, an eyepiece observation module and a control module according to functions. A spatial light modulator is adopted to load a pyramid phase, so that a Gaussian-Bessel illumination light field can be generated at the backfield of a focusing objective lens. In a detection optical path, a twisted Dammann grating is additionally adopted so as to realize simultaneous imaging of a multi-surface object, fluorescence collected from the objective lens simultaneously images a plurality of axial planes to an electron-enhanced CCD detection surface through a multi-surface imaging technology, and therefore, real-time three-dimensional fluorescence imaging can be realized. The real-time three-dimensional fluorescence imaging technique has an important practical value in biological living tissue and living cell microscopic imaging.

Description

Technical field [0001] The invention relates to a novel three-dimensional microscopic imaging technology, in particular to a real-time three-dimensional fluorescence microscopic imaging device. Background technique [0002] Optical imaging technology is a non-contact, non-destructive imaging technology, so it has important applications in imaging of living organisms or living cells. In addition to non-contact and non-destructive detection, in vivo biological imaging also requires real-time dynamic three-dimensional imaging information. At present, this kind of real-time 3D imaging technology is generally based on spatial scanning of galvanometers or piezoelectric ceramics or frequency domain scanning of broadband light sources. These dynamic 3D imaging technologies are dynamic imaging based on scanning technology, not real-time dynamic imaging in the true sense. In 1999, the British scientist Blanchard et al. [Appl. Opt. 38, 6692 (1999)] proposed a 4D imaging technology that si...

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Problems solved by technology

However, the twisted grating proposed by Blanchard et al. was originally designed based on the amplitude type grating, therefore, its main diffraction energy is concentrated in the zero order, and the overall energy efficiency is very low

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