Multi-Angle 4Pi Microscope Imaging System
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Solution Overview
Problem
Current microscopic imaging technologies face challenges in achieving high-resolution three-dimensional imaging while minimizing photobleaching and phototoxicity, and in obtaining high signal-to-noise ratios in fluorescence imaging.
Innovation Solution
A rapid three-dimensional imaging system and method based on a multi-angle 4Pi microscope, utilizing a spatial light modulator, wavefront modulation, and illumination interference to generate point spread functions with different inclination angles, enabling two-dimensional scanning and interference image acquisition for enhanced resolution and signal-to-noise ratio.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If light field microscopy technology is used to achieve real-time three-dimensional imaging, then imaging speed is improved, but spatial resolution deteriorates
Solution Approach 1:
The patent changes the illumination parameters by using structured light patterns (point, line, plane illumination modes) and adjusts the wavefront modulation parameters through the LCOS spatial light modulator. This allows the system to achieve high-resolution three-dimensional imaging by modifying the illumination characteristics rather than using conventional widefield illumination, thereby resolving the contradiction between imaging speed and spatial resolution
Solution Approach 2:
The patent introduces angular diversity by rotating the illumination pattern in different orientations and combining multiple illumination angles. This adds an angular dimension to the imaging process, enabling the system to capture three-dimensional information with high resolution while maintaining rapid imaging capability through the structured illumination approach
2Device complexity
If conventional fluorescence imaging is used, then imaging process is simple, but signal-to-noise ratio deteriorates due to photobleaching and phototoxicity
Solution Approach 1:
The patent employs periodic structured illumination patterns that are projected onto the sample in a time-varying manner. By using alternating illumination and dark periods, the system reduces the total fluorescence exposure time, thereby minimizing photobleaching and phototoxicity while maintaining high signal-to-noise ratio through the periodic modulation of the illumination
Solution Approach 2:
The patent applies structured illumination that selectively illuminates only specific regions or planes of the sample at any given time, rather than illuminating the entire sample uniformly. This localized illumination approach reduces the overall fluorescence signal exposure, decreasing photodamage while maintaining high signal-to-noise ratio in the imaged regions
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The system achieves high-resolution three-dimensional imaging with improved signal-to-noise ratio and expanded depth of field, enabling efficient imaging of living body samples by modulating the wavefront and polarization of light, and utilizing sparse collection and reconstruction algorithms.
Implementation Method 1
a wavefront modulation module, configured to place the LCOS device on a Fourier plane of an illumination end and to perform a wavefront modulation on the parallel light by projecting an intensity pattern
Implementation Method 2
an illumination interference module, configured to generate point spread function PSFs of a 4Pi through an illumination interference to irradiate a fluorescent sample
Implementation Method 3
acquire interference images of two fluorescent signals
Implementation Method 4
a controller, configured to control the wavefront modulation module to adjust a polarization direction of the light to generate PSFs of the 4Pi with different inclination angles
Data Source
AI summary
The present disclosure provides a rapid three-dimensional imaging system based on a multi-angle 4Pi microscope. The system includes: an illumination module, configured to obtain a parallel light of which a size covering a projection surface of a spatial light modulator; a wavefront modulation module, configured to place the LCOS device on a Fourier plane of an illumination end; a two-dimensional scanning module, configured to control a light beam to realize a two-dimensional scanning on an object plane; an illumination interference module, configured to generate point spread function PSFs of a 4Pi through an illumination interference to irradiate a fluorescent sample; an imaging module, configured to acquire interference images of two fluorescent signals; and a controller, configured to control the wavefront modulation module to adjust a polarization direction of the light to generate PSFs of the 4Pi with different inclination angles.