Light Sheet Microscopy for High-Resolution 3D Imaging
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Solution Overview
Problem
Current microscopes are inadequate for high-resolution, three-dimensional viewing of millimeter-sized biological objects due to issues with absorption, resolution loss, and limited ability to handle large samples or provide rapid data acquisition, especially in developmental biology applications.
Innovation Solution
A microscope design featuring a focusing arrangement that produces a two-dimensional object illumination pattern orthogonal to the detection beam path, allowing for relative movement between the illumination and sample, enabling high-resolution three-dimensional imaging with improved contrast and resolution by scanning narrow regions of the sample and using a mobile arrangement with a cylindrical lens or light mirrors for precise control of the light sheet.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a conventional light microscope is used for imaging millimeter-sized biological objects, then the sample can be viewed, but the resolution is lost and absorption occurs
Solution Approach 1:
The patent divides the illumination into a thin light sheet that scans across the sample, illuminating only a narrow plane at a time. This segmentation of the illumination volume allows high-resolution imaging of specific planes while minimizing overall light absorption and photodamage to the large millimeter-sized biological sample.
Solution Approach 2:
The patent transitions from conventional three-dimensional bulk illumination to a two-dimensional light sheet that is scanned through the sample. This dimensional change enables optical sectioning capability, allowing high-resolution imaging of specific planes within the thick sample while reducing absorption and photodamage compared to conventional volumetric illumination.
2Productivity
If the entire sample is illuminated simultaneously, then data acquisition is rapid, but neighboring fluorescence interferes with resolution
Solution Approach 1:
The illumination is segmented into a thin light sheet that scans across the sample, illuminating only a narrow plane at a time. This segmentation of the illumination volume allows high-resolution imaging of specific planes while minimizing overall light absorption and photodamage to the large millimeter-sized biological sample.
Solution Approach 2:
The patent transitions from conventional three-dimensional bulk illumination to a two-dimensional light sheet that is scanned through the sample. This dimensional change enables optical sectioning capability, allowing high-resolution imaging of specific planes within the thick sample while reducing absorption and photodamage compared to conventional volumetric illumination.
3Adaptability or versatility
If a cuvette is used as sample holder, then individual particles can be detected, but millimeter-size samples cannot be handled
Solution Approach 1:
The patent employs a dynamic scanning mechanism where the light sheet moves through the sample volume, allowing the system to handle large millimeter-sized samples by sequentially imaging different planes. This dynamic illumination approach replaces the static cuvette configuration, enabling versatile handling of various sample sizes while maintaining ease of operation through automated scanning.
4Measurement precision
If point illumination is used, then confocal resolution is achieved, but imaging time increases significantly
Solution Approach 1:
The illumination is segmented into a thin light sheet that scans across the sample, illuminating only a narrow plane at a time. This segmentation of the illumination volume allows high-resolution imaging of specific planes while minimizing overall light absorption and photodamage to the large millimeter-sized biological sample.
Solution Approach 2:
The patent transitions from conventional three-dimensional bulk illumination to a two-dimensional light sheet that is scanned through the sample. This dimensional change enables optical sectioning capability, allowing high-resolution imaging of specific planes within the thick sample while reducing absorption and photodamage compared to conventional volumetric illumination.
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
This design allows for high-resolution, three-dimensional imaging of millimeter-sized biological samples with improved contrast and reduced interference from neighboring fluorescence, enabling rapid data acquisition and efficient energy use, while minimizing sample stress and deformation.
Implementation Method 1
a focusing arrangement for producing a two-dimensional object illumination pattern
Implementation Method 2
using a laser and detects the fluorescent signals produced in the illumination light plane
Implementation Method 3
detects the fluorescent signals produced in the illumination light plane
Implementation Method 4
it does not have any means of moving the sample relative to the illumination light plane
Data Source
AI summary
The invention relates to a microscope in which a layer of the sample is illuminated by a plurality of thin strips of light (11) passed through a grid (34) and the sample is viewed (5) perpendicular to the plane of the strips of light. To record the image, the object (4) is displaced through the strips of light (11). At least three different images of the objects (4) are made at different phase angles. The images can be combined to form a single combined image.