Paraboloidal Mirror Light Disc Microscopy Shadowing Reduction
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Solution Overview
Problem
Current Light Sheet Fluorescence Microscopy (LSFM) systems face the challenge of 'shadowing' due to optically inconsistent light paths through the sample, leading to striping patterns that complicate the quantification of fluorescence intensity, which is not adequately addressed by existing solutions involving multiple co-planar light sheets that complicate alignment and have practical limits.
Innovation Solution
The introduction of a theoretically infinite number of coplanar light sheets using a single optical element, a paraboloidal mirror, to focus a disc of light on the sample, reducing shadowing to a minimum by ensuring every point is illuminated equally from all angles, compatible with any detection objective.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-affected harmful factors
If multiple co-planar light sheets are introduced to reduce shadowing, then shadowing is reduced, but device complexity increases due to complicated alignment and practical limits on the number of sheets
Solution Approach 1:
Multiple cylindrical lenses that would generate separate light sheets are merged into a single paraboloidal mirror that generates all light sheets simultaneously. This consolidation eliminates the alignment complexity inherent in using multiple separate optical elements while maintaining the shadow-reducing benefit of multiple illumination angles.
Solution Approach 2:
The paraboloidal mirror performs multiple functions that would otherwise require separate optical elements: it generates multiple coplanar light sheets at different angles, focuses them to a common focal plane, and eliminates shadowing artifacts. This single universal optical element replaces what would traditionally require multiple specialized components.
2Device complexity
If a single optical element is used to generate multiple coplanar light sheets, then device complexity is reduced, but manufacturing precision requirements increase for the paraboloidal mirror
Solution Approach 1:
The invention employs a paraboloidal (curved) mirror surface instead of flat or simple cylindrical surfaces. This specific curvature geometry inherently focuses light rays from different angles to a common focal plane, achieving the multiple light sheet function with a single element. The curved surface precision requirements are well-established in optical manufacturing and can be met using standard techniques.
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 the quality and accuracy of fluorescence microscopy images by minimizing shadowing, reducing out-of-focus fluorescence and photobleaching, and is applicable to both live-cell and fixed samples, maximizing image quality and quantitative information.
Implementation Method 1
directing a beam of annularly collimated excitation light on the mirror to focus a disc of light on the sample
Implementation Method 2
beam of annularly collimated excitation light
Implementation Method 3
Light Sheet Fluorescence Microscopy (LSFM) has emerged as a powerful approach to restrict the excitation volume of fluorescence microscopy
Data Source
AI summary
Methods for imaging a sample using fluorescence microscopy, systems for imaging a sample using fluorescence microscopy, and illumination systems for fluorescence microscopes. In some examples, a method includes positioning the sample such that a plane of interest of the sample is coplanar with a focal plane of a detection objective of a microscope; positioning a mirror around the sample; directing a beam of annularly collimated excitation light on the mirror to focus a disc of light on the sample; and imaging the sample through the detection objective.