Multi-Directional Light Sheet Microscopy Illumination
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Solution Overview
Problem
Microscopy techniques face challenges in achieving even illumination and rejecting out-of-focus light, particularly in highly scattering samples like biological tissues, where obstructions can block illumination, leading to shadowed regions and degraded image quality.
Innovation Solution
The multi-directional digital scanned light sheet microscopy (mDSLM) system employs an illumination sheet with higher angular diversity along one axis than the other, using beam shaping optics and scanning optics to ensure even illumination and reject out-of-focus light through a confocal aperture, allowing light to penetrate deeper and around obstructions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Illumination intensity
If conventional illumination is used to image a sample, then the field of view can be illuminated, but certain structures within the sample block the illumination light from reaching shadowed regions
Solution Approach 1:
The patent transforms conventional single-direction illumination into multi-directional illumination by introducing angular diversity along both the x and y axes. The illumination sheet is configured to propagate along the z-axis while having controlled angular diversity in orthogonal directions, allowing light to reach shadowed regions from multiple angles and eliminate occlusion artifacts.
Solution Approach 2:
The illumination sheet employs asymmetric angular diversity where the first angular diversity along the x-axis differs from the second angular diversity along the y-axis. This asymmetric configuration optimizes illumination penetration in different directions, allowing tailored illumination strategies for samples with specific geometric characteristics or obstruction patterns.
2Illumination intensity
If conventional illumination is used in scattering media, then light can reach the sample, but out of focus light scattered from outside the focal region reaches the detector
Solution Approach 1:
The illumination sheet provides highly localized illumination at the focal plane with minimal out-of-focus illumination. By confining the light sheet to the focal region and using controlled angular diversity, the system illuminates only the plane of interest while minimizing scattered light from above and below the focal plane, thereby improving signal-to-background ratio.
Solution Approach 2:
The system transitions from conventional omnidirectional or broad-angle illumination to a constrained light sheet geometry with specific angular diversity characteristics. This dimensional constraint in angular space ensures that only light from the focal region reaches the detector, effectively rejecting out-of-focus scattered light while maintaining penetration into scattering media.
3Length of stationary object
If a light sheet with high angular diversity is used, then light can penetrate deeper and reach occluded regions, but the system complexity increases
Solution Approach 1:
The patent achieves depth penetration and occlusion bypass by introducing angular diversity in the angular domain rather than physically moving the light source through large spatial distances. The illumination optics generate a light sheet with controlled angular spread along orthogonal axes, enabling deep penetration into scattering media and around obstructions while maintaining a compact system architecture.
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 quality by ensuring even illumination and reducing the impact of scattering and obstructions, enabling clearer imaging of samples with improved depth penetration and reduced blurriness.
Implementation Method 1
The beam shaping optics may include a diffraction grating. Each of the plurality of illumination beams may be a Gaussian pencil beam.
Implementation Method 2
The beam shaping optics may include a cylindrical telescope. The illumination sheet has a first angular diversity along a first axis and a second angular diversity along a second axis orthogonal to the first axis.
Implementation Method 3
The sample may include at least one fluorophore. The illumination sheet may excite the at least one fluorophore. The collection optics may receive emission light emitted by the excited ones of the at least one flourophores.
Data Source
AI summary
Embodiments disclosed herein directed to multi-direction digital light sheet microscopy (mDSLM). In mDSLM a sample may be illuminated with an illumination sheet which has a first angular diversity along a first axis and a second angular diversity along a second axis which is orthogonal to the first axis. An mDSLM system includes beam shaping optics which may passively reshape light into the illumination sheet. In some embodiments, scanning optics may be used to scan the illumination sheet with respect to the sample. In some embodiments, a confocal aperture may be used to limit out of focus light which reaches a detector of the mDSLM system.