Light Sheet Microscope Z-Stack Reconstruction
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Solution Overview
Problem
Current light sheet microscopes face challenges in reducing photonic sample exposure and increasing detection speed while maintaining high imaging quality.
Innovation Solution
A light sheet microscope with an adjustable light sheet plane and focal plane, allowing for step-by-step displacement through the sample volume, enabling the recording of a z-stack with reduced image numbers through overlap and mathematical reconstruction, thereby minimizing photonic exposure and enhancing recording speed.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a light sheet microscope records a complete z-stack with sequential light sheet positioning, then comprehensive sample coverage is achieved, but photonic exposure increases and detection speed decreases
Solution Approach 1:
The patent applies partial action by recording only a subset of images at different light sheet positions rather than complete sequential coverage. The selection criterion is that adjacent recorded images show an overlap of at least 50%, which is sufficient for mathematical reconstruction of intermediate images. This reduces the total number of images recorded while maintaining adequate sample coverage through the z-stack.
2Measurement precision
If a light sheet microscope records a complete z-stack with sequential light sheet positioning, then comprehensive sample coverage is achieved, but detection speed decreases
Solution Approach 1:
The patent applies partial action by recording only a subset of images at different light sheet positions rather than complete sequential coverage. The selection criterion is that adjacent recorded images show an overlap of at least 50%, which is sufficient for mathematical reconstruction of intermediate images. This reduces the total number of images recorded while maintaining adequate sample coverage through the z-stack.
3Object-affected harmful factors
If fewer images are recorded in the z-stack, then photonic exposure is reduced and detection speed increases, but image reconstruction quality may deteriorate
Solution Approach 1:
The patent applies feedback by implementing a quality assessment mechanism that evaluates whether the overlap between adjacent recorded images is sufficient (at least 50%). This feedback criterion ensures that the reduced set of recorded images maintains adequate information for mathematical reconstruction, preventing deterioration of reconstruction quality while still reducing photonic exposure compared to complete z-stack recording.
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
Significantly reduces photonic exposure of the sample and increases detection speed by up to three times while maintaining high imaging quality, with the ability to mathematically reconstruct intermediate images for improved sample representation.
Implementation Method 1
Owing to the illumination with the light sheet, only dyes of the sample in the light sheet plane that in embodiments corresponds to individual sample planes are excited in a targeted manner. This excitation generates fluorescent radiation from the light sheet plane.
Data Source
AI summary
In a light sheet microscope, a detection device images a sample volume and an illumination device illuminates the sample with a light sheet. A light sheet plane is arranged substantially perpendicularly to the optical imaging axis and an illumination width extends perpendicular to the light sheet plane. A processing device is provided to control setting of the location of the light sheet plane in the sample volume, and recording a plurality of images of the sample volume with different locations of the light sheet plane. For the plurality of images recorded, a spacing, which is measured along the optical imaging axis, between adjacent locations of the light sheet plane is greater than or equal to half the illumination width, but not greater than the illumination width. Intermediate images for the stack of images in additional locations of the light sheet plane in the sample volume are computed.