TDI Line-Scanning Microscopy With Multi-Slit Optical Sectioning
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Solution Overview
Problem
Traditional optical sectioning methods for TDI-based microscopy require expensive and bulky laser instrumentation, restrict scanning speed due to low fluorescence or luminescence signals, and can cause sample damage, limiting throughput and multi-color imaging capabilities.
Innovation Solution
A multi-slit or multi-pinhole illumination/detection system with a static mask in the intermediate image plane, allowing structured illumination and simultaneous detection of multiple color channels using TDI-based line scanning, with separate detectors for in-focus and out-of-focus signals.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional optical sectioning methods are used with single point/line focus illumination, then optical sectioning capability is achieved, but expensive and bulky laser instrumentation is required
Solution Approach 1:
The patent divides the illumination into multiple discrete slits or pinholes arranged in an array, replacing the traditional single point/line focus approach. This segmentation allows optical sectioning to be achieved through widefield fluorescence excitation with simpler illumination sources, eliminating the need for expensive laser instrumentation while maintaining sectioning capability.
Solution Approach 2:
The patent uses a mask with multiple slits or pinholes that creates multiple copies of the illumination pattern in the sample plane. This copying approach enables optical sectioning through structured illumination using simple widefield excitation sources rather than requiring complex single-point laser systems.
2Measurement precision
If single point/line focus illumination is used for optical sectioning, then sectioning is achieved, but scanning speed is restricted due to low fluorescence signal
Solution Approach 1:
By segmenting the illumination into multiple slits or pinholes, the patent increases the total illuminated area in the sample plane while maintaining optical sectioning. This allows more fluorophores to be excited simultaneously, generating stronger fluorescence signals that enable faster scanning speeds without sacrificing sectioning capability.
Solution Approach 2:
The patent merges multiple illumination points into an array of slits or pinholes, combining the excitation of multiple fluorophores simultaneously. This merging approach increases the overall fluorescence signal intensity, enabling faster scanning while maintaining optical sectioning through the mask-based structured illumination.
3Measurement precision
If high local intensities are used for optical sectioning, then sectioning is achieved, but sample damage may occur
Solution Approach 1:
The patent segments the illumination intensity distribution across multiple slits or pinholes rather than concentrating high intensity at a single point or line. This segmentation distributes the excitation energy over a larger area, reducing peak intensities and minimizing photodamage to the sample while maintaining optical sectioning capability through the structured illumination pattern.
4Adaptability or versatility
If multi-color imaging is implemented with traditional methods, then multiple color channels can be detected, but time-consuming iterative imaging is required
Solution Approach 1:
The patent implements a universal mask structure with multiple slits or pinholes that can simultaneously support multiple color channels. The mask design allows different wavelengths to pass through different regions, enabling simultaneous multi-color imaging without requiring iterative scanning or sequential acquisition, thus eliminating time loss while maintaining multi-color capability.
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
Enhances throughput and multi-color imaging capabilities while reducing cross-talk, enabling efficient optical sectioning and computational enhancement without the need for complex rotating elements, and supports high-resolution imaging of biological samples.
Implementation Method 1
Light detection through the same grid leads to efficient optical sectioning since mainly the light from in-focus radiation is transmitted through the grid and out-of-focus light is reflected by the non-transmissive regions of the mask
Implementation Method 2
out-of-focus light is reflected by the non-transmissive regions of the mask
Implementation Method 3
light from in-focus radiation is transmitted through the grid
Implementation Method 4
A continuous line scanning microscope based on time-delayed-integration (TDI) camera detection offers significantly higher throughput compared to traditional stop-and-stare approaches
Implementation Method 5
The TDI-based detection is therefore a favorable candidate for various existing imaging systems, as well as for the next generation biological imaging systems
Data Source
AI summary
An imaging system for imaging a biological sample or another sample containing fluorescent molecules may include an optical system with a light source emitting light, wherein the light is directed by the optical system to the sample via at least one plane being conjugate to the image plane. The optical source may have an extended radiation pattern, in other words, the radiation beam may have an extent in the x- and y-planes, rather than a point source. The extent of the illumination region in x and y may be based on and matched to the detector area onto which the radiation may be imaged, preferably a TDI detector. This novel system may include genomics, proteomics and transcriptomics work flows.