Multi-Focal Structured Illumination with Microlens-Pinhole Arrays
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Solution Overview
Problem
Conventional microscopy techniques face limitations in resolution and scanning speed, particularly in structured illumination microscopy (SIM) systems, which sacrifice temporal resolution for higher lateral resolution and are prone to shot noise, while confocal microscopy reduces fluorescence emission signal strength and scanning speed.
Innovation Solution
A multi-focal structured illumination microscopy (MSIM) system using hardware components like microlens and pinhole arrays generates multi-focal excitation patterns, blocks out-of-focus emissions, scales in-focus emissions, and sums them to produce high-resolution images without significant signal loss, employing hardware operations rather than software processing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If structured illumination microscopy (SIM) is used to achieve higher lateral resolution, then spatial resolution is improved, but temporal resolution deteriorates due to the time required to acquire multiple raw images
Solution Approach 1:
The patent divides a single light beam into multiple focal points using a microlens array, creating multiple excitation points simultaneously. This segmentation allows parallel acquisition of information from different spatial locations, effectively increasing scanning speed while maintaining resolution enhancement capabilities
Solution Approach 2:
The patent combines multiple focal points into a multi-focal pattern that illuminates the sample simultaneously. By merging multiple excitation points into a coordinated pattern, the system achieves both high spatial resolution and improved temporal resolution through parallel processing
2Measurement precision
If confocal microscopy uses a tightly closed pinhole to eliminate out-of-focus emission light, then optical resolution is improved, but fluorescence emission signal level deteriorates
Solution Approach 1:
The patent segments the detection process by using multiple pinholes corresponding to multiple focal points. Each pinhole collects light from its corresponding focal region, allowing efficient light collection while maintaining optical sectioning capability
Solution Approach 2:
The patent changes the pinhole configuration from a single tightly closed pinhole to multiple pinholes with optimized sizes and positions. This parameter change allows the system to maintain optical resolution while collecting more fluorescence signal through the combined aperture area of multiple pinholes
3Illumination intensity
If confocal microscopy aligns excitation beam with pinhole/detector perfectly, then light signal detection is improved, but system complexity increases due to alignment requirements
Solution Approach 1:
The patent implements self-alignment through the optical design where the microlens array and pinhole array are positioned in conjugate planes. The system automatically maintains alignment between excitation and detection paths through optical reciprocity, eliminating the need for complex manual alignment procedures
Solution Approach 2:
The patent uses asymmetric positioning of the microlens array relative to the pinhole array, with each lens optically coupled to a corresponding pinhole. This asymmetric but systematic arrangement simplifies alignment by creating fixed optical pathways that are inherently aligned through the optical design
4Adaptability or versatility
If SIM performs optical sectioning computationally, then processing flexibility is improved, but shot noise increases due to background fluorescence in thick samples
Solution Approach 1:
The patent extracts out-of-focus light physically using spatial pinholes before detection, removing the harmful background fluorescence that causes shot noise. This physical extraction complements computational processing and significantly reduces noise in thick samples
Solution Approach 2:
The patent performs optical sectioning as a preliminary action before detection by using the pinhole array to block out-of-focus light. This preliminary physical filtering reduces the noise burden on subsequent computational processing, improving signal-to-noise ratio in thick samples
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 MSIM system achieves high-resolution images at high scanning speeds with minimal signal loss, improving optical sectioning and reducing shot noise, suitable for thick samples, comparable to confocal microscopy performance.
Implementation Method 1
A multi-focal structured illumination microscopy (MSIM) system uses an array of detectors, such as pixels in a camera image, wherein each of the detectors in the array produces a separate confocal image
Implementation Method 2
the sample generates a plurality of fluorescent emissions resulting from each multi-focal pattern
Implementation Method 3
a spatial pinhole arrangement to eliminate out-of-focus emission light from specimens
Data Source
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AI summary
Various embodiments (300, 400, 500) for a multi-focal selective illumination microscopy (SIM) system for generating multi-focal patterns of a sample are disclosed. The embodiments (300, 400, 500) of the multi-focal SIM system perform a focusing, scaling and summing operation on each generated multi-focal pattern in a sequence of multi-focal patterns that completely scan the sample to produce a high resolution composite image.