Polarization-modulated fluorescence differential microscopy imaging method and device based on electro-optic modulation technology

Abstract

The invention discloses a polarization-modulated fluorescence differential microscopic imaging device and method based on electro-optical modulation technology, which includes an illumination system for generating excitation beams, a detection system for collecting fluorescent signals from samples, and a computer for control and signal processing. The illumination system includes: a laser, used to emit laser beams; an electro-optic modulation module, used to modulate the linearly polarized light to quickly switch between two different polarization states; a vortex half-wave plate, used to modulate the linearly polarized light into The radially polarized light and the angularly polarized light are focused by the objective lens to form a solid spot and a hollow spot for illuminating the sample. The invention realizes the rapid conversion of the solid light spot and the hollow light spot, and improves the imaging speed of the fluorescent differential microscopic imaging system to a large extent.

Description

technical field [0001] The invention belongs to the field of confocal microscopic imaging, and in particular relates to a method and device for realizing rapid phase modulation fluorescence differential microscopic imaging based on electro-optic modulation technology. Background technique [0002] Due to the limitation of the diffraction limit, the traditional far-field fluorescence microscopy technology is usually difficult to achieve a resolution lower than half the wavelength, so its application in the fields of biomedicine, nanotechnology, and materials is greatly limited. In order to break through this limitation, since the 1990s, relevant researchers have proposed many super-resolution microscopy techniques such as stimulated radiation depletion super-resolution microscopy (STED), single-molecule localization super-resolution microscopy (SMS), etc. microtechnology. Among these super-resolution microscopy techniques, fluorescence differential microscopy (FED), as a rec...

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Problems solved by technology

Since the sample to be tested needs to be scanned with solid and hollow spots during the imaging process, when using a vortex phase plate as a phase modulation method, a beam splitter is required to split the beam into two paths, and one path is modulated into a hollow spot through the vortex phase plate. The other beam is not phase-modulated into a solid spot, and the sample to be tested is scanned with two beams, so that the complexity of the system and the difficulty of adjustment will be relatively high; when using a spatial light modulator as a phase modulation method, usually by changing the spatial light modulator The phase modulation pattern loaded on the surface of the modulator realizes the conversion between solid and hollow spots. Since the spatial light modulation itself has a certain response time, and the phase modulation pattern loaded by the spatial light modulator is controlled by a software program to switch between the two modes. It also takes a certain amount of time to switch between solid and hollow spots, so it takes a certain amount of time to switch between solid and hollow spots. It will take a long time to scan the sample to be tested with a solid spot and a hollow spot, and motion artifacts will appear, which will limit the fluorescence. Imaging speed of differential microscopic imaging and observation of moving samples

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