Collimated Microscopic Imaging via Parallel Line Scanning
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Solution Overview
Problem
Existing microscopic imaging techniques using nonlinear sample interactions for enhanced spatial resolution face limitations due to sequential data acquisition, reduced fluorescence emission, and technical complexities in achieving parallel data acquisition while maintaining confocal imaging and resolution.
Innovation Solution
A method and arrangement utilizing line-shaped excitation and confocal detection with a line camera behind a slit diaphragm, enabling parallel data acquisition by structuring the illumination with coherent light diffraction and interference, allowing for simultaneous excitation of multiple spots with axially structured switching light, and using phase masks to optimize light distribution for high-resolution imaging.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If sequential data acquisition is used with point scanning methods, then confocal imaging quality is maintained, but image compilation time increases significantly
Solution Approach 1:
The patent divides the imaging process into multiple parallel scanning paths that can be processed simultaneously. By segmenting the data acquisition into independent linear scan segments that can be handled in parallel, the system maintains confocal imaging quality while significantly reducing total image compilation time through concurrent processing of multiple segments.
Solution Approach 2:
The patent transitions from sequential point-by-point scanning to parallel linear scanning by adding a spatial dimension. Instead of scanning one point at a time, the system scans multiple linear paths simultaneously, effectively moving from one-dimensional sequential acquisition to multi-dimensional parallel acquisition, thereby reducing time loss while maintaining imaging quality.
2Measurement precision
If resolution is increased fivefold laterally, then spatial precision improves, but fluorescence emission decreases twenty-five times
Solution Approach 1:
The patent applies preliminary action by pre-switching dyes to the switched-off state before the actual fluorescence measurement. This allows the system to achieve high spatial resolution through nonlinear optical interactions while maintaining sufficient fluorescence emission, as the dye is already in the appropriate state for both high resolution and adequate signal generation.
Solution Approach 2:
The patent changes the optical parameters by using nonlinear optical interactions and switching dyes between different states (on/off). This allows the system to achieve fivefold lateral resolution improvement while maintaining acceptable fluorescence emission levels, as the parameter changes enable efficient energy utilization at the reduced excitation volumes.
3Productivity
If multi-spot arrangement is used for parallelization, then data acquisition speed improves, but confocal detection becomes cumbersome and resolution enhancement is limited
Solution Approach 1:
The patent segments the detection system into multiple independent linear scan detectors that can operate in parallel. Each detector handles a specific linear scan path, allowing simultaneous data acquisition across multiple spots without requiring complex confocal detection arrangements. This segmentation simplifies the overall system while maintaining high productivity.
Solution Approach 2:
The patent uses a line camera to capture multiple linear scan paths simultaneously, adding a spatial dimension to the detection process. Instead of complex multi-spot confocal detection, the system uses a simplified linear scanning approach that can be parallelized across multiple detectors, reducing device complexity while improving data acquisition speed.
4Measurement precision
If doughnut-shaped illumination is used for nonlinear interaction, then resolution enhancement is achieved, but the area interacting with light increases reducing parallelization density
Solution Approach 1:
The patent applies local quality by using line-shaped illumination instead of doughnut-shaped illumination. The line shape provides the necessary nonlinear interaction for resolution enhancement while concentrating the light into a thinner, more localized region. This allows higher parallelization density as multiple line spots can be packed more closely together compared to doughnut spots.
Solution Approach 2:
The patent changes the illumination geometry from two-dimensional doughnut shapes to one-dimensional line shapes. This dimensional change allows for more efficient packing of illuminated spots, increasing parallelization density while maintaining the nonlinear optical interactions necessary for resolution enhancement through the line-shaped illumination pattern.
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 high-resolution imaging beyond diffraction limits with efficient parallel data acquisition, reducing image compilation times and maintaining confocal imaging quality, thereby overcoming the limitations of sequential data acquisition and technical complexities in multi-spot arrangements.
Implementation Method 1
structuring the illumination with coherent light diffraction and interference
Implementation Method 2
structuring the illumination with coherent light diffraction and interference
Implementation Method 3
enabling parallel data acquisition by structuring the illumination with coherent light diffraction and interference, allowing for simultaneous excitation of multiple spots
Implementation Method 4
using phase masks to optimize light distribution for high-resolution imaging
Data Source
AI summary
A method and arrangement for collimated microscopic imaging, including a first illumination of a sample in at least one region for exciting fluorescence, and a spatially resolving detection of the sample light by detector elements, the detection being associated with the region, wherein by means of a second illumination a sub-division of the region into separate fluorescent partial regions occurs, which are associated with the detector elements. The separation of the partial regions is carried out by the spatial separation of the fluorescent regions by means of intermediate regions having reduced fluorescence or no fluorescence, and/or by means of different spectral properties of the fluorescence from the partial regions.