Angled reflective surface for transverse illumination microscopy
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Solution Overview
Problem
Current microscopy techniques face limitations in achieving high-resolution 3D imaging of live samples due to optical resolution limits, complex instrumentation, and invasive illumination methods, particularly in Selective Plane Illumination Microscopy (SPIM), which requires specialized machinery and sample holders, making it unsuitable for routine laboratory use.
Innovation Solution
A sample holding device with angled reflective surfaces that allows transverse illumination of a sample using a single objective, enabling 3D SPIM imaging with high and super-resolution capabilities on standard microscopes, facilitating imaging of single cells to whole organisms without moving the sample, and allowing simultaneous imaging of multiple cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional light microscopy is used, then the setup is simple and suitable for routine laboratory use, but the resolution is limited to above 250 nm laterally and 500 nm axially
Solution Approach 1:
The patent segments the illumination and detection functions by using separate optical paths - illumination objectives positioned at angles (e.g., 45 degrees) to the detection objective, allowing transverse illumination while maintaining simple detection geometry compatible with routine microscopes
Solution Approach 2:
The patent makes standard microscope objectives multi-functional by using them both for detection and for creating the light sheet through transverse illumination, eliminating the need for specialized SPIM objectives while achieving super-resolution
2Measurement precision
If STED, SIM, or single-molecule techniques are used to achieve ultra-high resolution, then spatial resolution is improved, but deep imaging capabilities in live biological samples are reduced and the techniques require complex instrumentation and data analysis
Solution Approach 1:
The patent extracts the illumination function from the detection path by implementing transverse illumination through angled objectives, separating the light sheet creation from the fluorescence collection path to simplify instrumentation while maintaining super-resolution capabilities
Solution Approach 2:
The patent inverts the traditional epifluorescence configuration by illuminating the sample from the side (transverse illumination) rather than from above through the same objective used for detection, enabling light sheet microscopy with standard objectives
3Object-affected harmful factors
If Fluorescence Light-Sheet microscopy is used, then imaging of live cells is improved with minimal invasiveness, but complex machinery and difficult set ups make this method unsuitable for routine laboratory practise
Solution Approach 1:
The patent makes standard microscope objectives multi-functional by using them for both detection and transverse illumination, eliminating the need for specialized SPIM objectives and complex sample holders, thereby simplifying the system for routine laboratory use
Solution Approach 2:
The patent inverts the conventional microscopy configuration by positioning illumination objectives at angles to the detection objective, creating transverse illumination that enables light sheet microscopy with standard inverted microscopes and regular coverslips
4Measurement precision
If SPIM requires 2 objectives placed perpendicularly with special sample holders, then light sheet microscopy is achieved, but high NA objectives and regular coverslips cannot be used
Solution Approach 1:
The patent inverts the traditional SPIM configuration by using transverse illumination through angled objectives rather than perpendicular illumination, allowing the use of standard inverted microscopes, high NA objectives, and regular coverslips
Solution Approach 2:
The patent segments the optical paths by positioning illumination and detection objectives at different angles (e.g., 45 degrees separation), allowing independent optimization of each path and compatibility with standard microscope components
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
The device provides excellent 3D optical sectioning with acquisition speeds comparable to spinning-disc microscopy, enabling high-resolution imaging of large samples and reducing acquisition time, while being compatible with standard microscopes and allowing for the imaging of multiple cells simultaneously.
Implementation Method 1
one or more sides of the well are provided with an angled reflective surface adapted to reflect a light sheet transversely through a sample
Data Source
Figure 1A~1C
Figure 2Ai~2Eiii
Figure 3Ai~3Bvi
AI summary
A sample holding device for use in transverse illumination of a sample or sub-components of a sample comprising: a support substrate comprising a sample well adapted to contain and be compatible with said sample wherein said well is provided on at least one wall with an angled reflective surface adjacent said sample well which when in use directs a transverse light beam from a light source through a sample contained within said sample well to provide substantially transverse illumination of a sample contained therein and imaging the sample using a single objective.