High-Throughput Hyperspectral Imaging With Snapshot Depth Sectioning
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Solution Overview
Problem
Hyperspectral imaging systems face challenges such as bleaching due to tunable filters and limited throughput in data acquisition, especially when dealing with thick tissue samples, and confocality issues in wide-field approaches.
Innovation Solution
A high-throughput hyperspectral imaging system utilizing a combination of excitation light sources, lenslet arrays, and channel separators to simultaneously capture multiple spatially dispersed spectral channels, enabling four- or five-dimensional imaging with depth sectioning and reduced spectral distortion.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If tunable filters are used to capture fluorescence at high spectral resolution, then spectral resolution is improved, but bleaching occurs and emission intensity changes during scanning
Solution Approach 1:
The patent replaces the mechanical scanning system with a snapshot imaging system that uses a diffractive optical element to spatially disperse wavelengths across a two-dimensional detector array. This eliminates the need for sequential scanning through spectral bands, thereby preventing bleaching-related intensity changes while maintaining high spectral resolution through optical dispersion.
Solution Approach 2:
The patent transitions from one-dimensional spectral scanning to two-dimensional snapshot imaging by using a diffractive optical element to map wavelengths spatially across the detector plane. This dimensional change allows simultaneous capture of the entire spectrum without sequential scanning, resolving the contradiction between spectral resolution and intensity stability.
2Area of stationary object
If wide-field approach is used to capture fluorescence, then field of view is improved, but confocality and depth sectioning are lost
Solution Approach 1:
The patent introduces a spatial light modulator that segments the wide-field fluorescence into multiple focal planes, enabling confocal detection at different depths simultaneously. This segmentation allows the system to maintain both wide field of view and depth sectioning capability by processing different depth layers in parallel.
3Measurement precision
If scanning is used to acquire hyperspectral data cube, then depth sectioning is achieved, but data acquisition speed is limited
Solution Approach 1:
The patent implements continuous parallel acquisition by using a snapshot imaging system that captures the entire hyperspectral data cube in a single shot. The diffractive optical element continuously disperses wavelengths across the detector while the spatial light modulator maintains confocal detection across multiple depths simultaneously, eliminating the time loss associated with sequential scanning.
4Measurement precision
If patterned illumination is scanned across the sample, then depth sectioning is achieved, but throughput is limited
Solution Approach 1:
The patent uses a spatial light modulator to project patterned illumination across multiple depth planes simultaneously in the axial dimension, while the snapshot detector captures the entire three-dimensional dataset in a single exposure. This dimensional approach to parallelization maintains depth sectioning capability while achieving high throughput by eliminating temporal sequencing.
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 throughput and reduced bleaching by simultaneously acquiring hyperspectral data in multiple dimensions, allowing for efficient analysis of biological samples, including thick tissues, with improved confocality and reduced spectral distortion.
Implementation Method 1
Each of the plurality of first lenslet arrays is configured to receive light from the light source and to generate a pattern of light
Implementation Method 2
The objective is configured to simultaneously image each of the patterns of light to form a plurality of parallel lines or an array of circular spots at different depths of the sample
Implementation Method 3
the excitation light causes the sample to emit fluorescence light
Implementation Method 4
a channel separator that is configured to receive the fluorescence light from the sample and separate the fluorescence light into a plurality of spatially dispersed spectral channels
Data Source
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AI summary
High-throughput hyperspectral imaging systems (100) are provided. According to an aspect of the invention, a system includes an excitation light source (110); an objective (140) that is configured to image excitation light onto the sample (150), such that the excitation light causes the sample to emit fluorescence light; a channel separator (170) that is configured to separate the fluorescence light into a plurality of spatially dispersed spectral channels; and a sensor (180). The excitation light source (110) includes a light source and a plurality of lenslet arrays. Each of the lenslet arrays is configured to receive light from the light source and to generate a pattern of light, and the patterns of light generated by the lenslet arrays are combined to form the excitation light. The objective is configured to simultaneously image each of the patterns of light to form a plurality of parallel lines or an array of circular spots at different depths of the sample.