Microscope Image-Guided Illumination for High-Speed Axial Precision
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Solution Overview
Problem
Existing systems lack the capability to perform high-content, high-speed image-guided microscopic illumination for processes such as photobleaching, photoactivation, and light-triggered reactions in specific subcellular areas, with techniques like STOMP and LCM lacking axial precision and speed for efficient biomolecular sample collection.
Innovation Solution
A microscope-based system integrating optical, photochemical, and mechatronic designs, utilizing a processing module to control imaging and illumination assemblies for real-time image processing and pattern illumination, enabling rapid image-guided illumination at 300 milliseconds per field of view, with a femtosecond laser for two-photon precision.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If manual STOMP technique is used, then axial precision can be achieved, but processing speed and high-content capability are insufficient
Solution Approach 1:
The patent replaces manual mechanical operation with automated image processing and computer-controlled illumination systems. Image processing algorithms automatically identify regions of interest, and computer-controlled scanners deliver precise illumination, eliminating manual intervention while achieving both high axial precision and high processing speed.
Solution Approach 2:
The patent employs two-photon fluorescence microscopy with femtosecond laser pulses to achieve precise axial localization. By changing the illumination parameters (using nonlinear optical processes and precise pulse timing), the system achieves both high axial precision and rapid processing speeds unattainable with conventional single-photon systems.
2Ease of operation
If LCM system is used, then tissue isolation capability is provided, but axial precision and speed for biomolecular sampling are insufficient
Solution Approach 1:
The patent replaces mechanical laser cutting with optical two-photon fluorescence excitation for biomolecular sampling. This substitution enables precise axial localization at the subcellular level while maintaining ease of operation through automated image-guided illumination, overcoming the limitations of conventional LCM systems.
Solution Approach 2:
The patent extends isolation capability from the two-dimensional plane (LCM) to three-dimensional space by utilizing the optical sectioning capability of two-photon fluorescence microscopy. This enables precise sampling at specific depths within tissues while maintaining automated operation and high speed.
3Productivity
If high-speed illumination is implemented, then processing throughput increases, but illumination precision and control accuracy may deteriorate
Solution Approach 1:
The patent implements real-time feedback through image processing that continuously monitors and adjusts illumination positioning. The system processes images to identify regions of interest and automatically adjusts scanner positioning and illumination timing, ensuring high precision is maintained even at high processing throughput speeds.
Solution Approach 2:
The patent employs synchronized periodic action where illumination pulses are precisely timed with image acquisition cycles. By coordinating the scanning mirrors and laser pulses with the image processing timing, the system achieves both high throughput and high precision through rhythmic, synchronized operation.
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
Enables high-content processing of proteins, lipids, and nucleic acids for proteomic, transcriptomic, and metabolomic studies by rapidly collecting enough biomolecular samples within a reasonable duration, achieving high axial illumination precision and efficiency.
Implementation Method 1
A femtosecond laser may be used as the illumination light source to generate a two-photon effect for high axial illumination precision
Data Source
AI summary
A system and method for image-guided microscopic illumination are provided. A processing module controls an imaging assembly such that a camera acquires an image or images of a sample in multiple fields of view, and the image or images are automatically transmitted to a processing module and processed by the first processing module automatically in real-time based on a predefined criterion so as to determine coordinate information of an interested region in each field of view. The processing module also controls an illuminating assembly to illuminate the interested region of the sample according to the received coordinate information regarding to the interested region, with the illumination patterns changing among the fields of view.