Microscope Image Splitting for Fast Low-Stress 3D Imaging
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Solution Overview
Problem
Existing microscopes face limitations in achieving high image and volume rates while minimizing sample stress and maintaining manageable equipment complexity, particularly in three-dimensional imaging of biological samples, due to inefficiencies in light exposure and inflexible confocal diaphragm arrangements.
Innovation Solution
A microscope design that splits emission light into multiple partial beam paths, generating partial images on the camera sensor surface, synchronized with the position of excitation light on the sample, allowing for simultaneous readout of these images and reducing sample irradiation time.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If line illumination is used to achieve high frame rates, then higher volume rates are possible, but sample stress increases due to high radiation intensity over extended axial area
Solution Approach 1:
The detection of emission light is segmented into multiple axial planes using movable reflective elements (confocal diaphragms) positioned at different depths. Each reflective element captures emission from a specific axial plane and directs it to the camera, enabling parallel detection from multiple depths without increasing illumination intensity at any single plane.
Solution Approach 2:
The system transitions from detecting emission from a single axial plane to simultaneously detecting emission from multiple axial planes by adding the axial dimension to the detection process. This is achieved by positioning reflective elements at different axial positions to capture and redirect light from conjugate planes at different depths to the camera sensor.
2Productivity
If multiple reflective elements are used to detect emission from multiple axial planes, then volumetric rate increases, but device complexity increases due to fixed and immutably integrated beam path
Solution Approach 1:
The confocal diaphragms are implemented as movable reflective elements that can be dynamically adjusted along the optical axis. This mobility allows the system to adapt the detection planes to different imaging requirements and objectives, transforming a static complex system into a dynamic one that can reconfigure itself for different measurement tasks.
Solution Approach 2:
The movable reflective elements serve multiple functions: they act as confocal diaphragms for optical sectioning, as beam directors to guide light from different axial planes to the camera, and as adjustable components that can be positioned according to the specific imaging requirements. This multi-functionality reduces the need for separate dedicated components for each function.
3Measurement precision
If aperture sizes are fixed at one Airy unit, then confocal imaging is achieved, but adaptability to different resolution levels and image field sizes is limited
Solution Approach 1:
The aperture sizes of the confocal diaphragms are made dynamically adjustable rather than fixed at one Airy unit. This allows the system to adapt the aperture size according to the specific imaging requirements, such as changing resolution levels or field sizes, while maintaining confocal imaging capabilities. The adjustability enables optimization of the balance between resolution and light throughput for different measurement tasks.
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 approach enhances image and volume rates with reduced sample stress and equipment complexity, enabling faster and gentler volumetric scanning with lower light exposure.
Implementation Method 1
an illumination objective for illuminating and scanning a sample with excitation light... a microscope objective for directing emission light emitted by the sample
Data Source
Figure 1
Figure 2~3
AI summary
The invention relates to a microscope having: an illumination beam path with an illumination control device for illuminating a specimen; a detection beam path for directing radiated emission light toward a camera and having a control unit for controlling the illumination control device and the camera, wherein the control unit is designed to synchronise regions to be read of a sensor surface of the camera with a position of the excitation light, which position is defined by the illumination control device; and having an image splitter unit in the detection beam path for splitting the emission light into a plurality of partial beam paths which each produce a partial image of the specimen on the sensor surface of the camera, wherein the partial images lie next to one another such that linear regions in the partial images, which regions correspond to a position of the excitation light defined by the illumination control device on or in the sample, lie on the same line or the same lines of the sensor surface.