Slit-Scan Microscopy for Large-Sample Imaging Without Stitching
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Solution Overview
Problem
Existing microscopy methods struggle to efficiently capture images of samples larger than the field of view without significant time and resolution loss, particularly in applications like screening on microtiter plates, due to the need for multiple stop&go cycles and stitching of images.
Innovation Solution
A method involving a continuously moving sample with a slit-shaped illumination synchronized with a detector, allowing for image acquisition without stitching by controlling the relative movement between the sample and microscope, and correcting for image deformations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Area of stationary object
If multiple images are captured by moving the sample to map areas larger than the field of view, then the coverage area is increased, but the acquisition time increases significantly due to multiple stop&go cycles
Solution Approach 1:
The patent implements continuous scanning of the sample through the field of view without stopping, replacing the conventional stop&go approach. The detector continuously records signal as the sample moves, enabling seamless coverage of large areas. This continuous action eliminates the time loss associated with repeated acceleration, deceleration, and positioning operations, directly resolving the contradiction between coverage area and acquisition time.
Solution Approach 2:
The patent segments the large sample area into multiple scanning passes or trajectories. Instead of capturing the entire large area in a single static frame, the system divides the imaging task into sequential segments along the scanning path. Each segment is captured continuously as the sample moves through the field of view, and the segments are later reconstructed into a complete image of the entire sample area.
2Area of stationary object
If multiple images are captured and stitched together to cover large areas, then the coverage area is increased, but the manufacturing complexity increases due to image stitching operations
Solution Approach 1:
The patent incorporates reference markers or fiducial features directly into the sample or mounting substrate before imaging. These pre-placed references enable automatic alignment and registration during the stitching process. By performing this preparatory action beforehand, the complex image stitching operation is simplified, as the software can automatically match and align images based on the known reference positions, reducing the overall processing complexity.
3Measurement precision
If high magnification is used to capture detailed images, then the resolution is improved, but the field of view decreases requiring more images to cover large areas
Solution Approach 1:
The patent transitions from a two-dimensional static imaging approach to a three-dimensional approach by adding the time dimension through continuous scanning. Instead of attempting to capture the entire large area at high magnification in a single frame (which is physically impossible), the system captures a narrow strip of high-resolution data continuously as the sample moves through the field of view. This effectively adds the scanning direction as a third dimension, allowing high resolution in the detected dimensions while covering large areas through the scanning motion.
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 high-resolution imaging of large sample areas with minimal resolution loss and reduced acquisition time, achieving up to √2-fold resolution enhancement in the scan direction and maintaining resolution in orthogonal directions.
Implementation Method 1
illuminating, by a light source controlled by the at least one control unit, a part of the sample through the microscope objective
Implementation Method 2
detecting, by a detector controlled by the at least one control unit, light from the sample collected by the microscope objective
Data Source
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AI summary
A method for acquiring microscope-images of sample being larger than a field of view of a microscope, the method comprising creating a continuous relative movement between a sample and the microscope, wherein the optical axis of a microscope objective is substantially perpendicular to the vector of the relative movement, illuminating a part of the sample through the microscope objective, wherein the illuminated part of the sample is smaller than the field of view and forms an illumination slit, moving the illumination slit in a scanning direction across the field of view, and detecting light from the sample collected by the microscope objective, wherein the sample is moved in the same direction as the scanning direction or in a direction perpendicular to the scanning direction while the illumination slit is moved across the field of view.