Cell Plaque Detection Using Multi-Focus Bright-Field Imaging
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Solution Overview
Problem
Current cell culture imaging systems require laborious maintenance, stringent aseptic conditions, and are not efficient in distinguishing between live and lysed cells, especially in plaque detection, necessitating improved imaging methods for cell analysis.
Innovation Solution
The use of Phase Field in focus and non-focused images to detect cell objects, combined with the Transport of Intensity Equation methodology, allows for the detection of live cells and segmentation of lysed cell materials, utilizing bright field optics and image processing techniques to create a Phase Gradient image.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If bright field optics are used to image cells, then the imaging system is simple and cost-effective, but the contrast between cells and surrounding media is very low making it difficult to distinguish live cells from lysed cell materials
Solution Approach 1:
The patent segments the imaging process into multiple focal planes (above focus and below focus) to extract different optical characteristics. By analyzing images at different z-positions, the system separates live cell signals from lysed cell debris through phase behavior analysis, resolving the contrast problem while maintaining bright field optics simplicity
Solution Approach 2:
The patent introduces the z-dimension (focal plane depth) to the imaging analysis. By capturing images at multiple focal planes and analyzing phase differences along the z-axis, the system creates additional discrimination dimensions that enable accurate distinction between live cells and lysed materials without adding complex optical components
2Productivity
If conventional imaging methods are used, then the imaging process is fast, but the system cannot reliably detect plaque regions and distinguish between normal cells and lysed cell materials
Solution Approach 1:
The patent performs preliminary imaging at multiple focal planes before final analysis. By capturing and processing images at both above-focus and below-focus planes in advance, the system pre-extracts phase behavior characteristics that enable reliable plaque detection in subsequent analysis, maintaining imaging speed while improving detection reliability
Solution Approach 2:
The patent uses phase behavior as an intermediary characteristic to bridge the gap between raw imaging data and plaque detection. The phase differences captured at different focal planes serve as an intermediate representation that enables reliable distinction between live cells and lysed materials, improving detection accuracy without sacrificing imaging speed
3Measurement precision
If phase contrast optics are used to improve cell visibility, then cell contrast and detection accuracy improve, but the system complexity and cost increase
Solution Approach 1:
The patent creates a computational copy of phase contrast imaging by processing bright field images at multiple focal planes. Through image processing algorithms that analyze phase behavior across z-positions, the system generates phase-like information from simple bright field optics, achieving phase contrast capabilities without the complex optical hardware
Solution Approach 2:
The patent replaces the mechanical/optical phase contrast system with a computational approach. Instead of using complex phase contrast optics to physically separate and enhance cell contrast, the system uses digital image processing of bright field images to computationally extract phase behavior, substituting optical complexity with algorithmic processing
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 accurate detection and segmentation of live and lysed cells, providing objective data for cell culture monitoring, improving repeatability, traceability, and enabling quantitative measures of cell growth and morphology, with applications in cell incubators and standalone imaging systems.
Implementation Method 1
Cells are composed of material that differs from the surrounding media mainly in the refractive index. This results in very low contrast when the cells are imaged with bright field optics. Phase contrast optics utilizes the different phase delay of the inner material and the surrounding media.
Implementation Method 2
Live cells with an organized shape concentrate the illumination, forming bright spots in the above focus regions of the field. This concentration of illumination also creates a virtual darkened region in the field below the in-focus plane.
Implementation Method 3
This behavior is the phenomena behind the Transport of Intensity Equation methodology for recovering the phase of the bright field illuminated subjects.
Data Source
AI summary
A plaque detection method and apparatus wherein at least one processor is programmed to receive above focus images to detect the presence of live cells without detecting the lysed cell materials, receive below focus images wherein virtual dark regions exist which are similar to cell shadows as seeds in a segmentation process and use contours around each resulting shape to obtain a subset that are more likely to be part of the cell population to define a cell map. A distance map is created in which each pixel value is the distance of that pixel from the nearest pixel of the cell map and the distance map is thresholded to create a first image of the places which are relatively far from the cells a second image with a smaller distance threshold to get an image that mimics the edges of the cells.