Common-Path Parallel Fluorescence Emission Super-Resolution Microscopy
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Solution Overview
Problem
Existing parallel fluorescence emission difference microscopy (pFED) is limited by external disturbances such as drift and noise due to separate excitation light paths, which degrade imaging quality.
Innovation Solution
Implementing a common-path parallel fluorescence emission difference microscopy method using a liquid crystal spatial light modulator to modulate excitation light paths, ensuring they share the same optical path and are affected equally by disturbances, allowing for improved signal-to-noise ratio and imaging speed through staggered solid and doughnut-shaped spots on the sample plane.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If parallel fluorescence emission difference microscopy uses two separate excitation light paths to achieve faster imaging speed, then imaging speed is improved, but imaging quality deteriorates due to external disturbances such as drift and noise
Solution Approach 1:
The patent merges the two separate excitation light paths into a common-path configuration, where both the solid spot and doughnut-shaped spot share the same optical path from the objective lens to the sample. This eliminates differential drift and noise between paths, resolving the contradiction by maintaining high imaging quality while preserving the parallel scanning capability for fast imaging speed.
Solution Approach 2:
The patent introduces a spatial light modulator as an intermediary device that generates both the solid spot and doughnut-shaped spot from a single excitation light path. This intermediary enables the creation of two excitation patterns without requiring separate optical paths, thus maintaining imaging quality while achieving parallel scanning for fast imaging speed.
2Measurement precision
If conventional fluorescence emission difference microscopy uses sequential scanning to maintain high imaging quality, then imaging quality is preserved, but imaging speed deteriorates
Solution Approach 1:
The patent segments the single excitation light path into two parallel paths using a spatial light modulator, generating both the solid spot and doughnut-shaped spot simultaneously. This segmentation enables parallel scanning of the sample, achieving fast imaging speed while maintaining high imaging quality through the common-path configuration.
Solution Approach 2:
The patent implements continuous parallel scanning by generating both excitation patterns simultaneously and scanning them across the sample at the same time. This eliminates the sequential operation, achieving continuous useful action that provides both fast imaging speed and high imaging quality.
3Productivity
If separate excitation light paths are used to enable parallel scanning, then imaging speed is improved, but reliability deteriorates due to sensitivity to external disturbances
Solution Approach 1:
The patent merges the excitation paths into a common-path configuration, making both the solid spot and doughnut-shaped spot susceptible to the same environmental disturbances. This merging eliminates differential drift and noise, resolving the contradiction by maintaining reliability while preserving the parallel scanning capability for high imaging speed.
Solution Approach 2:
The patent applies local quality modification by using a spatial light modulator to create different spot patterns (solid and doughnut-shaped) at specific locations in the optical path. This local modification enables parallel scanning for fast imaging speed while the common optical path maintains robustness to drift and noise.
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 method enhances imaging quality by canceling out disturbances, improving the signal-to-noise ratio and enabling faster super-resolution imaging with increased detail and resolution.
Implementation Method 1
A polarization component of the linearly polarized light parallel to a direction which can be modulated by the spatial light modulator is modulated using half of the liquid crystal spatial light modulator by 0-2πvortex phase modulation
Implementation Method 2
liquid crystal spatial light modulator
Implementation Method 3
Two paths of light emitted from the other half of the spatial light modulator is converted into circularly polarized light
Implementation Method 4
the laser beam is converted into linearly polarized light by a polarizer
Implementation Method 5
The other half of the liquid crystal spatial light modulator is loaded as a blazed grating, such that a unmodulated component of the linearly polarized light is modulated to be inclined
Implementation Method 6
the solid spot and the doughnut-shaped spot scanning a sample at the same time, and excited two paths of fluorescence signals pass through respective detection light paths and is received by two detectors
Data Source
AI summary
A super-resolution microscopic imaging method and apparatus based on common-path parallel fluorescence emission difference microscopy. In the method, a liquid crystal spatial light modulator is used to modulate excitation light in fluorescence emission difference microscopy super-resolution microscopic imaging, and two parts of the spatial light modulator are respectively loaded into 0-2π vortex phase modulation and blazed grating, so that the common-path excitation light forms solid spot and doughnut-shaped spot with a certain distance on a sample surface at the same time, and parallel scanning is carried out, thereby ensuring that the imaging speed is doubled compared with that of ordinary fluorescence emission difference super-resolution microscopic imaging, and at the same time, the two excitation lights are not easily affected by noise, drift and other interferences due to the common path.