Multiphoton subpulse STED-SPIM microscopic system realized by single wavelength

Abstract

The invention relates to a multiphoton subpulse STED-SPIM (Selective Plane Illumination Microscopy) microscopic system realized by single wavelength, characterized in that the femtosecond pulse laser emitted by a femtosecond single wavelength laser is transmitted to a first splitter through a first polarization regulator to be divided into two beams of light, and a first beam of light is successively transmitted to a second polarization regulator, a second splitter, a third polarization regulator and a quarter-wave plate along a Y-axial direction to form an excitation light beam; a second beam of light is transmitted to an STED photoquenching system, and the light outputted by the STED photoquenching system is successively emitted into the second splitter, the third polarization regulator and the quarter-wave plate to be modulated into donut-shaped focus spots to form a quenching light beam; a scanner is used for scanning the excitation light beam and the quenching light beam to generate an excitation plate light source and an STED quenching plate light source; an excitation plate light source and an STED quenching plate irradiate an object to be imaged through an excitation objective lens to allow the flourescent dye of the object to be imaged to excite fluorescence, and excited fluorescence is imaged through an imaging objective lens, and is emitted to a photosensitive element or an analysis element successively through an optical filter and a convergent lens.

Description

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AI Technical Summary

Benefits of technology

This patented innovation allows multiple photons (called pulses) from one source to create an image called Multiplex Imaging (MI). These images have improved sensitivity compared to traditional methods such as Time Division Multiple Access (TDMA), Spatial Harmonic Analysis (SHARI), Lasers Direct Structured Illumination Microscopy (SDLIM), etc., while reducing costs associated therewith. Additionally, this new technique improves efficiency when implementing Single Wavelength Exposure Light Pulse Interference Compensated Fluorescences (SWIFLF)) imaging techniques. Overall these improvements improve overall performance and make it possible to combine them together without requiring expensive equipment like Optically Stabilized Lifetime Contrast Focal Microscope (CSMLCM)/Superlenses Acquisition(SSA)).

Problems solved by technology

This patented technical problem addressed in this patents relates to improving the performance of multiplexed plasmon microscopes while reducing their own limitations such as limited spatial resolution and reduced sensitivity compared to current techniques like stochastic emission depletion microscpy.

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