Object Imaging with Asymmetric Light Modulation for Focus Detection
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Solution Overview
Problem
Current image cytometry and microscopy methods face challenges in accurately identifying and segmenting biological particles, require complex optical components, and struggle with determining the focus and position of objects along the optical axis, especially under difficult conditions or when using birefringent materials.
Innovation Solution
A method involving the collection and modulation of transmitted, scattered, or emitted light to form multiple images with asymmetric modulation and different focal planes, allowing for the processing of these images to enhance contrast and determine object position without changing optical components, using techniques like phase-contrast and bright-field imaging.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional microscopy methods are used to depict biological particles, then the basic imaging function is achieved, but the contrast and ability to resolve signal intensities is insufficient
Solution Approach 1:
The patent segments the imaging process into multiple channels: transmitted light imaging for background and position, scattered light imaging for contrast enhancement, and fluorescence imaging for specific molecular detection. Each channel processes different aspects of the sample independently, allowing optimal detection parameters for each while simplifying the overall optical design compared to attempting single-channel optimization.
Solution Approach 2:
The imaging system is designed with multi-functional capability to perform transmitted light imaging, scattered light imaging, and fluorescence imaging using a shared optical platform. This universal approach allows the same system to achieve high contrast across different imaging modes without requiring separate specialized optical paths for each technique.
2Measurement precision
If fluorescence microscopy is used to achieve high contrast, then the signal-to-background ratio is improved, but the method becomes labour intensive and requires specialised training
Solution Approach 1:
The system employs automated image acquisition and processing algorithms that automatically adjust imaging parameters, segment cells, and analyze features without requiring manual intervention or specialized operator skills. The automated focus determination and multi-channel image processing occur independently, making the system easy to operate while maintaining high measurement precision.
3Loss of information
If focus stacks are recorded to determine spatial position in 3 dimensions, then the positional information is obtained, but the computational treatment is intensive and complicated
Solution Approach 1:
The patent extracts focus and positional information directly from single-plane images using automated algorithms that determine the focal plane and object positions without requiring acquisition of multiple focus stacks. This extraction approach obtains the necessary spatial information while avoiding the computational burden of processing multi-plane focus stacks.
Solution Approach 2:
The system replaces the mechanical approach of physically moving through multiple focal planes (focus stacking) with an computational algorithm that determines focus and position from a single plane image. This substitution eliminates the need for complex multi-plane image acquisition and processing while preserving the essential spatial information.
4Measurement precision
If fluorescent molecules are bound to objects for detection, then the contrast is improved, but the binding method is very selective and only a certain fraction of objects can give rise to a signal
Solution Approach 1:
The patent merges multiple detection approaches: transmitted light imaging provides universal background and position information for all objects, scattered light imaging enhances contrast for objects with different refractive indices, and fluorescence imaging provides specific molecular detection. This combination ensures that all objects are detected and characterized, with each method compensating for the limitations of the others.
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 provides optimal performance in various applications, maximizes fluorescence excitation/emission channels, and accurately determines object position and focus with minimal complexity, offering improved visualization and masking of biological objects.
Implementation Method 1
collecting transmitted, scattered, diffracted, or emitted light from the sample
Implementation Method 2
collecting transmitted, scattered, diffracted, or emitted light from the sample
Implementation Method 3
collecting transmitted, scattered, diffracted, or emitted light from the sample
Implementation Method 4
in emission microscopy, such as fluorescence microscopy, where light emitted onto a particle gives rise to emission of light at a higher wavelength
Implementation Method 5
for at least the first and the second image the collected light is modulated asymmetrically differently
Data Source
AI summary
A method for characterizing object(s) in a sample includes collecting transmitted, refracted, scattered, diffracted, and/or emitted light from the sample. The collected light formed on a sensor surface includes at least a first and second image of the object(s). For at least the first and second image, the collected light is modulated asymmetrically differently, or for at least a third and fourth image focal plane positions are different, or for at least a fifth image the collected light is modulated in at least two places differently compared to the surroundings, or for at least a sixth and seventh image of the object(s), the collected light is modulated asymmetrically differently and focal planes of the sixth and seventh image have different positions. The images are processed to characterize the objects(s).