Multi-Plane Microscope Using Segmented Illumination
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Solution Overview
Problem
Existing microscopy techniques struggle to efficiently image extended sample volumes without causing fluorophore saturation and often require complex setups to capture multiple sample planes.
Innovation Solution
A microscope design that simultaneously illuminates and spatially separates multiple sample regions using parallel offset illumination light beams, with separate detection channels to avoid crosstalk and reduce image saturation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If a single light sheet illuminates the sample plane, then the illumination is simple and the setup is straightforward, but the imaging speed is limited and fluorophore saturation occurs
Solution Approach 1:
The illumination light beam is divided into multiple parallel offset light beams that simultaneously illuminate different sample planes. This segmentation allows concurrent imaging of multiple planes, increasing imaging speed while avoiding fluorophore saturation by distributing the illumination across separate planes rather than concentrating it in a single plane.
Solution Approach 2:
The patent transitions from illuminating a single sample plane to illuminating multiple sample planes simultaneously by adding the dimension of parallel offset illumination. This dimensional expansion enables volumetric imaging capability, improving productivity without requiring complex scanning mechanisms.
2Productivity
If multiple sample planes are imaged simultaneously, then imaging efficiency increases, but crosstalk between detection channels occurs
Solution Approach 1:
The detection system is segmented into multiple separate detection channels, each dedicated to detecting fluorescence from a specific sample plane. This segmentation prevents crosstalk between planes by ensuring that each plane's signal is captured by its own dedicated channel, maintaining image quality while enabling simultaneous multi-plane imaging.
Solution Approach 2:
Each detection channel is optimized for its specific sample plane, with local quality control ensuring that fluorescence signals from different planes are distinguished and detected independently. This local optimization prevents signal interference and maintains high image quality across all imaged planes.
3Device complexity
If a single objective lens is used for both illumination and detection, then the setup is simplified, but the ability to spatially separate multiple sample planes is limited
Solution Approach 1:
A single objective lens is designed to perform multiple functions: it serves as both the illumination objective and the detection objective. The lens is configured to accept multiple parallel offset illumination beams and simultaneously focus fluorescence from multiple sample planes onto corresponding detection channels. This multi-functionality enables simplified setup while maintaining multi-plane imaging 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 efficient and gentle volume imaging by preventing fluorophore saturation and allowing for simultaneous imaging of multiple sample planes, thereby improving image contrast and reducing complexity.
Implementation Method 1
the sample is illuminated with a light sheet... The fluorescent light to be detected is coupled out between the lens and the scanning unit
Implementation Method 2
a scanning unit for scanning the sample with the illumination light... comprising a scanning mirror
Implementation Method 3
optics for focusing the illumination light emitted by the illumination unit into the sample and for focusing the detection light originating from the sample onto the detector
Data Source
Figure 1~2
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Figure 5~6
AI summary
The invention relates to a microscope (50) for imaging a sample (30), comprising an illumination unit (12) for emitting illumination light onto the sample (30), a detector (71) for detecting detection light from the sample (30), an optical unit for focusing the illumination light emitted by the illumination unit (12) into the sample (30) and for focusing the detection light from the sample (30) on the detector (71), and a scanning unit (20) for scanning the sample (30) with the illumination light. The illumination unit (12) is designed to emit illumination light in the form of separate illumination light bundles (52, 54, 56) on the scanning unit (20) in such a manner that the illumination light bundles (52, 54, 56) are focused simultaneously on spatially separate, strip-shaped sample regions (A, B, C) during the scanning of the sample (30). The detector (71) is designed to detect the detection light simultaneously and in a spatially separate manner in the form of separate detection light bundles (58, 60, 62) which stem from the spatially separate, strip-shaped sample regions (A, B, C).