Spectrally Selective Diffractive Element for Wavelength-Resolved Superresolution Microscopy
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Solution Overview
Problem
Current fluorescence microscopy systems face challenges with high costs, large installation space, and complex alignment requirements due to the need for multiple color channels and cameras, which also result in increased data processing and computational load, especially when achieving high-resolution imaging beyond the optical diffraction limit.
Innovation Solution
A fluorescence microscopy method and system that uses a spectrally selective diffractive element to generate diffraction images of isolated fluorescence labels onto a single detector, allowing for localization precision exceeding optical resolution and wavelength identification without the need for multiple color channels, using emitter pairs or ensembles to encode color information in diffraction patterns.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple color channels and cameras are used for wavelength-resolving imaging, then wavelength resolution is improved, but device complexity and installation space increase
Solution Approach 1:
The patent combines multiple wavelength channels into a single camera system by using a diffractive optical element that spatially separates different wavelengths. This merges the function of multiple cameras into one while maintaining wavelength resolution through diffraction-based spectral separation.
Solution Approach 2:
The diffractive optical element acts as an intermediary between the fluorescence labels and the detector. It mediates the wavelength information by diffracting different wavelengths to different spatial positions, enabling a single camera to resolve multiple wavelengths without requiring multiple separate detection channels.
2Measurement precision
If multiple cameras are used for high-resolution imaging, then imaging quality is improved, but alignment requirements and operational complexity increase
Solution Approach 1:
The patent merges multiple camera functions into a single detector system. By using a diffractive optical element to spatially separate wavelengths, all wavelength information can be captured on one camera, eliminating the need for precise alignment between multiple cameras while maintaining high imaging quality.
3Measurement precision
If computational methods are used to achieve superresolution, then spatial resolution is improved, but data processing load increases
Solution Approach 1:
The diffractive optical element serves as an intermediary that performs wavelength separation in the optical domain before detection. This physical separation reduces the computational burden by converting what would be complex computational deconvolution problems into simpler spatial pattern recognition tasks on the detector.
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 reduces the complexity and cost of the system, minimizes data processing, and eliminates the need for precise alignment of multiple cameras, while maintaining high-resolution imaging and wavelength resolution, thereby improving efficiency and flexibility in imaging biological samples.
Implementation Method 1
the imaging beam path of the microscope has a spectrally selective element of pre-defined diffraction properties, which element, during the imaging, diffracts the image of the sample comprising the isolated fluorescing labels into a first diffraction order
Implementation Method 2
fluorescence labels are repeatedly excited to emit fluorescence radiation
Data Source
AI summary
A method for wavelength-resolving and high spatial resolution fluorescence microscopy in which fluorescence labels in a sample are repeatedly excited to emit fluorescence radiation and frames including images of isolated labels are produced with a microscope. The positions of the images of the isolated fluorescing labels are localized with a localization precision exceeding the optical resolution of the imaging beam path of the microscope. The imaging beam path of the microscope has a diffractive element which, during the imaging, diffracts the image of the sample comprising the isolated fluorescing labels into a first diffraction order so that each frame contains the first diffraction order images of the isolated fluorescing labels. A parameter of the first diffraction order images of the isolated fluorescing labels is evaluated and an indication of the wavelength of the isolated fluorescing labels is derived from this evaluated parameter.