Multispot Scanning Microscopy with Independent Spectral Control
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Solution Overview
Problem
Current multispot scanning microscopy techniques face issues such as significant photo bleaching and damage to specimens, slow image recording, and increased noise due to the sequential scanning method, which limits image speed, signal-to-noise ratio, and flexibility in spectral excitation and detection.
Innovation Solution
A device and method that allow independent setting of the spectral composition of illumination and detection sub-beams using controllable beam manipulation means and spectral selection means, enabling optimized scanning speed, reduced photo damage, and improved signal-to-noise ratio, along with flexible spectral selection for various measurement tasks.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If sequential raster-scanning with a single laser point is used, then optical sectioning and blur-free imaging are achieved, but image recording speed is slow and photo damage increases
Solution Approach 1:
The patent divides the single scanning beam into multiple parallel scanning beams that simultaneously scan different regions of the specimen. This segmentation allows parallel image acquisition across multiple spots, dramatically increasing recording speed while maintaining optical sectioning quality through confocal detection for each beam
Solution Approach 2:
The patent combines multiple scanning beams into a single optical path using beam combining optics, allowing simultaneous illumination of multiple specimen regions. The detected signals from all beams are merged and processed together, enabling fast parallel imaging while preserving the optical sectioning capability of each individual beam
2Measurement precision
If high laser power is used in the focal spot for signal generation, then signal-to-noise ratio is improved, but photo bleaching and photo damage increase
Solution Approach 1:
The patent segments the total illumination power across multiple scanning beams, so each beam delivers lower power to its focal spot compared to a single high-power beam. This reduces photo damage and bleaching at each location while maintaining adequate signal-to-noise ratio through the combined signal from multiple beams
Solution Approach 2:
The patent implements continuous scanning motion for all beams simultaneously, ensuring that no single location receives prolonged high-power exposure. The continuous movement distributes the total energy load across the entire scanned area over time, reducing peak power density and associated photo damage
3Device complexity
If spectral composition is fixed for all illumination beams, then device complexity is reduced, but adaptability for different measurement tasks and fluorescence dyes is limited
Solution Approach 1:
The patent provides independent spectral control for each illumination beam through separate beam manipulation means, allowing different wavelengths and spectral compositions for each beam. This enables tailored excitation for different fluorescence dyes in different regions while maintaining manageable complexity through modular optical components
Solution Approach 2:
The patent implements dynamically adjustable spectral parameters for each beam that can be changed during operation. The beam manipulation means allow real-time modification of wavelength, bandwidth, and spectral shape to adapt to different measurement requirements and dye characteristics without physical reconfiguration
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 solution enables faster scanning, reduced photo damage, and enhanced signal-to-noise ratio, allowing for more precise and flexible imaging of specimens with improved spectral selectivity and compatibility with different fluorescence dyes.
Implementation Method 1
a splitting device for splitting the illumination light beam into a plurality of illumination sub-beams
Implementation Method 2
detection light radiated by the specimen in detection sub-beams after irradiation with the individual illumination sub-beams
Data Source
AI summary
The invention relates to a device for multispot scanning microscopy, having a multicolor light source for providing at least one illumination light beam, having a splitting device for splitting the illumination light beam into a plurality of illumination sub-beams, having first optical means for providing an illumination optical path for guiding and focusing the individual illumination sub-beams respectively into a light spot on or in a specimen to be examined, having a scan unit for guiding the light spots over the probe, having a detection unit for detecting detection light emitted by the specimen in detection sub-beams after irradiation with the individual illumination sub-beams, having second optical means for providing a detection optical path for guiding the detection sub-beams to the detector unit, having a control and evaluation unit for controlling the scan unit and for evaluating the detection light detected by the detection unit. The device is characterized in that in the illumination optical path for at least two of the illumination sub-beams a controllable beam manipulation means is present for independent setting of a spectral composition of the respective illumination sub-beam, and the control and evaluation unit is designed to control the beam manipulation means. The invention further relates to a method for multispot scanning microscopy.