Microscope Illumination Using Segmented Light Sources
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Solution Overview
Problem
Current microscopy techniques, such as MINFLUX and STED microscopy, face challenges with rapid and precise positioning of illumination light, especially with electro-optical scanners that are complex, costly, and inefficient, and have limited deflection angles, making it difficult to achieve high-resolution imaging below the diffraction limit.
Innovation Solution
A method and device for sequential point-shaped illumination using a predefined illumination point pattern with individual light sources, where each illumination point is illuminated with a focused light bundle and time-offset, allowing for flexible and rapid adaptation to sample-specific requirements, eliminating the need for complex scanners and achieving precise localization of fluorescent markers.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If electro-optical scanners are used for rapid positioning of illumination light, then positioning speed is improved, but device complexity and cost increase
Solution Approach 1:
The illumination system is divided into multiple independent light sources, each capable of illuminating specific illumination points simultaneously. This segmentation eliminates the need for a single complex scanner that would need to rapidly switch between points, instead using simpler, dedicated light sources for each point.
Solution Approach 2:
The patent transitions from temporal multiplexing (scanning points sequentially in time) to spatial multiplexing (illuminating multiple points simultaneously in space). By adding the dimension of multiple independent light sources, the system achieves rapid positioning without complex scanning mechanisms.
2Speed
If electro-optical scanners are used for rapid positioning, then positioning speed is improved, but power dissipation increases
Solution Approach 1:
By segmenting the illumination into multiple independent light sources, each source operates at lower power levels continuously rather than requiring high power bursts during rapid scanning transitions, reducing overall power dissipation while maintaining positioning speed.
3Device complexity
If a single illumination light source is used for sequential illumination, then device complexity is reduced, but illumination speed decreases
Solution Approach 1:
The single light source is segmented into multiple independent light sources, each dedicated to specific illumination points. This allows parallel illumination of multiple points simultaneously, dramatically increasing illumination speed while keeping each individual light source simple.
Solution Approach 2:
The system adds the dimension of multiple simultaneous light sources rather than relying on temporal sequencing. This spatial parallelization enables illumination speed to increase without increasing the complexity of individual light source components.
4Speed
If electro-optical scanners are used for positioning, then positioning speed is improved, but the maximum achievable deflection angle is limited
Solution Approach 1:
Instead of relying on a scanner to achieve large deflection angles through mechanical rotation, the patent uses multiple light sources positioned at different locations. This spatial arrangement allows effective illumination across a large field of view without being constrained by the maximum deflection angle of a single scanner.
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 enables precise localization of fluorescent markers with improved resolution and accuracy, reducing the need for costly and inefficient scanners, and allows for flexible illumination patterns, enhancing the quality of imaging and localization in microscopy.
Implementation Method 1
each of the individual light sources emits an illumination light beam that is focused onto its assigned illumination point by means of imaging optics
Implementation Method 2
As long as the fluorescent marker is within the doughnut ring, but not exactly at its zero point, it emits fluorescence photons that can be detected
Data Source
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AI summary
The invention relates to a method for illuminating a sample (1) in a microscope, in particular a MINFLUX microscope, in points using illumination light, wherein the sample (1) is illuminated sequentially at the illumination points (3) of a specified or specifiable illumination point pattern (2). The method is characterized in that a lateral extension of the illumination point pattern (2) is smaller than the longest wavelength of the illumination light, the illumination points (3) are illuminated constantly solely in a temporally offset manner, each illumination point (3) of the illumination point pattern (2) is assigned a dedicated individual light source of a plurality of individual light sources (4), and each illumination point (3) is illuminated using the focal point of an illumination light bundle (5) of the individual light source (4) assigned to the illumination point.