Image Scanning Microscope with Split Spatial-Spectral Detection
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Solution Overview
Problem
Current image scanning microscopes face limitations in quantifying the spectral content of emitted fluorescence light, leading to reduced usability and light loss due to higher diffraction orders and the use of a single polarization state.
Innovation Solution
An image scanning microscope with a detection arrangement that splits detection light into two parts, using a spatially resolved detector for intensity and a spectrally resolved detector for spectral information, enabling the determination of a fluorophore species' spatial distribution with high spatial resolution and signal-to-noise ratio.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a multi-element photodetector is used in ISM to improve spatial resolution and signal-to-noise ratio, then spatial resolution and signal-to-noise ratio are improved, but the ability to quantify spectral content is limited
Solution Approach 1:
The detection light is segmented into two separate beam paths using a beam splitting element. The first beam path directs light to a spatially resolved detector for high-resolution spatial imaging, while the second beam path directs light to a spectrally resolved detector for spectral content analysis. This segmentation allows both spatial and spectral information to be captured independently without compromising either measurement quality.
2Loss of information
If existing spectral detection solutions are used, then spectral information can be obtained, but light loss occurs due to higher diffraction orders and single polarization state limitation
Solution Approach 1:
The second beam path is designed to serve multiple functions: it captures spectral information across a broad wavelength range, handles multiple polarization states simultaneously, and minimizes diffraction-related light loss. The spectrally resolved detector in this path is configured to efficiently detect fluorescence across different wavelengths without the limitations of previous single-function designs, thereby reducing overall light loss while maintaining comprehensive spectral coverage.
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
Enables robust differentiation of multiple fluorophore species in a sample by combining spatial and spectral information, enhancing imaging capabilities beyond traditional confocal microscopy.
Implementation Method 1
a beam splitting element configured to receive the detection light, to split the detection light into two parts, to direct a first part of the detection light into a first beam path, and to direct a second part of the detection light into a second beam path
Implementation Method 2
The first beam path comprises a spatially resolved detector configured to detect a spatial distribution of an intensity of the first part of the detection light
Implementation Method 3
The second beam path comprises a spectrally resolved detector configured to detect a spectral information of the second part of the detection light
Implementation Method 4
Each photodetector element in the array is configured to output a detector signal upon receiving fluorescent light emitted from the sample
Data Source
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AI summary
An image scanning microscope (100, 200) comprises a detection arrangement (112, 204) comprising a beam splitting element (122) configured to receive detection light (120), to split the detection light (120) into two parts, to direct a first part (124a) of the detection light (120) into a first beam path (126a), and to direct a second part (124b) of the detection light (120) into a second beam path (126b). The first beam path (126a) comprises a spatially resolved detector (128, 202) configured to detect a spatial distribution of the intensity of the first part (124a) of the detection light (120). The second beam path (126b) comprises a spectrally resolved detector (130) configured to detect a spectral information of the second part (124b) of the detection light (120). The image scanning microscope (100, 200) further comprises a controller (116) configured to determine a spatial distribution of at least one fluorophore species in a sample (104) based on the spatial distribution of the intensity of the first part (124a) of the detection light (120) and the spectral information of the second part (124b) of the detection light (120).