Multi-spectral Scanning Microscopy with Sequential Detector Mixing
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Solution Overview
Problem
Existing high-resolution scanning microscopy methods require significant adjustment and are costly due to the need for multiple, expensive area detectors for color analysis, which reduces spatial resolution and increases complexity with multiple color channels.
Innovation Solution
A method and microscope design that uses a spatially resolving detector with spectral splitting and mixing, allowing radiation from different spatial and spectral channels to be merged into mixed channels, which are then directed to a single detector element, enabling multi-spectral imaging without reducing spatial resolution and simplifying adjustments across color channels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple area detectors are used for color analysis in high-resolution scanning microscopy, then spectral resolution is improved, but spatial resolution deteriorates and device complexity increases
Solution Approach 1:
The detection process is segmented into two sequential stages: first, spatial resolution is achieved using a single area detector to capture the diffraction image; second, spectral resolution is achieved by sequentially measuring fluorescence at different wavelengths at each spatial position. This segmentation allows both high spatial and spectral resolution without requiring multiple simultaneous detectors.
Solution Approach 2:
The spectral measurement is performed through periodic action by sequentially scanning different wavelengths at each spatial position. The system measures fluorescence intensity at multiple discrete wavelengths in sequence, creating a periodic measurement cycle that builds up the complete spectral information over time, rather than requiring all spectral channels to be captured simultaneously.
2Measurement precision
If multiple area detectors are used for color analysis, then spectral resolution is improved, but device complexity and cost increase
Solution Approach 1:
A single area detector is made multi-functional by sequentially using it for both spatial imaging and spectral measurement. The same detector hardware performs both functions at different time points, eliminating the need for multiple specialized detectors. The system achieves spectral analysis capability through wavelength-sequential measurement using the universal detector.
Solution Approach 2:
The single area detector serves itself for both spatial and spectral measurement tasks. Rather than requiring external additional detectors, the system uses the existing detector's full capability set by varying the measurement parameters (wavelength selection) to achieve both imaging and spectroscopy functions with one device.
3Loss of information
If multiple color channels are measured simultaneously, then spectral information is improved, but adjustment complexity increases
Solution Approach 1:
Instead of simultaneous multi-channel measurement requiring complex alignment, the system uses periodic sequential measurement at different wavelengths. Each wavelength is measured in turn using the same optical path and detector, which greatly simplifies adjustment and alignment requirements while still capturing complete spectral information through the time-sequential approach.
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 allows for high-resolution, multi-spectral imaging with reduced adjustment complexity and cost, maintaining spatial resolution while enabling color analysis across multiple channels without the need for multiple, expensive detectors.
Implementation Method 1
a method for high-resolution scanning microscopy of a sample wherein the sample is excited by illumination radiation to emit fluorescence radiation
Data Source
AI summary
For the purposes of high-resolution scanning microscopy, a sample is excited by illumination radiation to emit fluorescent radiation in such a way that the illumination radiation is focused at a point in or on the sample, so as to form a diffraction-limited illumination spot. The point is imaged in a diffraction image on a detector in a diffraction-limited manner, wherein the detector has detector elements and a plurality of location channels which resolve a diffraction structure of the diffraction image. The sample is scanned with various scanning positions with an increment smaller than half the diameter of the illumination spot. An image of the sample with a resolution that is increased beyond a resolution limit of the image is generated from the data of the detector and from the scanning positions associated with these data. In order to distinguish between at least two predetermined spectral channels in the fluorescent radiation of the sample, for each location channel there is an independent beam path leading to a separating element that spectrally divides these beam paths into the spectral channels and then remixes the spectral channels of the different location channels into the same number, on additional independent beam paths, such that a plurality of the additional independent beam paths receive the radiation in different spectral channels and from different location channels, and each of these additional independent beam paths leads to one of the detector elements.