Light Sheet Microscope Alignment Using Real-Time Pupil Image Feedback
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Solution Overview
Problem
Existing methods for adjusting light sheet alignment in microscopes require stopping the microscope operation for parameter optimization, which is not suitable for continuous data acquisition.
Innovation Solution
A system and method that uses a beam splitter to intercept a fraction of light, forms two bidimensional images on a photodetector, and employs a data processing unit to calculate misalignment parameters in real time, allowing continuous alignment adjustment without stopping image acquisition.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If parameter optimization based on merit figure linked to contrast is performed, then alignment accuracy is improved, but data acquisition speed deteriorates due to stopping microscope operation
Solution Approach 1:
The system enables continuous alignment adjustment during microscope operation by processing images in real-time. The beam splitter continuously monitors light sheet alignment while the microscope acquires data, eliminating the need to stop operation for optimization. This maintains both high alignment accuracy and continuous data acquisition.
Solution Approach 2:
The system implements a feedback mechanism where the beam splitter continuously measures light sheet alignment by comparing images from different focal planes. The data processing unit calculates misalignment parameters and generates adjustment commands that are fed back to the alignment system, enabling real-time correction without interrupting data acquisition.
2Measurement precision
If manual periodic alignment adjustments are performed, then alignment accuracy is improved, but time loss due to stopping operation increases
Solution Approach 1:
The system performs self-alignment by automatically monitoring and adjusting light sheet position using the beam splitter and data processing unit. The microscope service itself (alignment adjustment) is performed by the system without external intervention, eliminating the need for manual periodic adjustments and associated time losses.
Solution Approach 2:
Alignment adjustment becomes a continuous automated process rather than periodic manual intervention. The system continuously monitors alignment parameters and makes adjustments as needed, eliminating downtime and maintaining both accuracy and productivity.
3Difficulty of detecting and measuring
If beam splitter is introduced to intercept light fraction, then alignment monitoring capability is improved, but light loss increases
Solution Approach 1:
The beam splitter extracts only a small fraction of the light for alignment monitoring purposes, while the majority of light continues to the detector for image acquisition. This selective extraction enables alignment monitoring with minimal impact on the main imaging function and light availability.
Solution Approach 2:
The beam splitter is positioned to intercept only the necessary fraction of light for alignment monitoring, concentrating the monitoring function in a specific location without affecting the overall light path for imaging. This local intervention minimizes light loss while maintaining monitoring capability.
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 continuous alignment adjustment of the light sheet while the microscope operates, maintaining high image quality and reducing the need for manual periodic alignments.
Implementation Method 1
a beam splitter configured to intercept a fraction of light emitted by the slice of the sample
Implementation Method 2
an arrangement of optical elements and a photodetector, the arrangement of optical elements focusing the intercepted fraction of light emitted by the sample slice to form two bidimensional images of the sample slice on the photodetector
Data Source
AI summary
It is disclosed a system and method for adjusting the alignment of a light sheet microscope. A beam splitter intercepts a fraction of light emitted by a slice of a sample illuminated by the light sheet. The intercepted light fraction is focused to form two bidimensional images on a photodetector, each bidimensional image being formed by rays coming from a respective portion of an exit pupil of the microscope's objective. At least two pairs of corresponding areas are identified in the two bidimensional images, and a mutual distance between corresponding areas of each pair is determined. A misalignment parameter indicative of a misalignment of the light sheet relative to a focal plane of the microscope's objective is then calculated, based on the determined mutual distances of the various pairs of corresponding areas. An adjustment command is then generated based on the misalignment parameter.