Microscopy Image Modulation for Contrast and 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 setups, and struggle with limited fluorescence channels and focus determination without altering sample conditions or changing optical components.
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 enhanced contrast and focus determination without mechanical movement of optical components.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Illumination intensity
If fluorescent microscopy is used to achieve high contrast, then contrast is improved, but the method becomes labor intensive and requires specialized training
Solution Approach 1:
The patent replaces manual fluorescent staining procedures with automated image processing techniques. Instead of requiring laboratory technicians to perform fluorescent microscopy steps (which are labor intensive and require specialized training), the system uses computational methods to enhance contrast and identify particles automatically, substituting mechanical/lab procedures with digital processing
Solution Approach 2:
The patent creates multiple synthetic images of the same sample by applying different modulation patterns and processing algorithms. These virtual copies allow the system to extract contrast information without physically altering the sample or requiring complex optical setups, enabling automated analysis without specialized training
2Measurement precision
If focus stacks are recorded to determine 3D spatial position, then measurement precision is improved, but computational complexity increases
Solution Approach 1:
The patent extracts focus information from individual 2D images without requiring full focus stack processing. By using asymmetric modulation patterns and analyzing contrast changes in single images, the system determines z-axis position and depth of field without computing complex focus stacks, thereby reducing computational requirements while maintaining measurement precision
Solution Approach 2:
The patent segments the imaging process into independent contrast enhancement steps applied to individual images. Instead of processing a complete focus stack (multiple images at different focal planes), the system analyzes each image separately using asymmetric modulation, dividing the complex 3D reconstruction problem into simpler 2D contrast analysis tasks
3Adaptability or versatility
If multiple fluorescent dyes are used to detect different objects, then adaptability is improved, but the number of available channels is limited
Solution Approach 1:
The patent changes the detection parameter from wavelength (fluorescence channels) to spatial frequency (modulation patterns). Instead of using multiple fluorescent dyes with different emission wavelengths (which are limited in number), the system applies multiple asymmetric modulation patterns to the same light path, creating distinguishable contrast patterns that can be computationally separated without being limited by the number of spectral channels
4Measurement precision
If optical components are moved to determine focus, then measurement precision is improved, but device complexity increases
Solution Approach 1:
The patent replaces mechanical focus adjustment mechanisms with computational focus determination. Instead of physically moving optical components (objectives, lenses, or detectors) to different focal planes to determine focus, the system uses asymmetric modulation patterns and image processing algorithms to calculate focus status and z-axis position from stationary 2D images, eliminating mechanical movement while maintaining precision
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 precise characterization of biological objects with improved contrast and z-axis elevation, optimizing performance across various microscopy applications without component changes, and facilitating faster focus determination.
Implementation Method 1
for at least the first and the second image the collected light is modulated asymmetrically differently
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
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
at least a third image and a fourth image of the object(s), wherein focal planes of the third image and of the fourth image have different positions
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).