Cell Plaque Detection Using Phase Field Bright-Field Imaging
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Solution Overview
Problem
Current cell culture imaging systems require laborious processes, stringent aseptic conditions, and are not efficient in distinguishing between live and lysed cells, especially in plaque detection, necessitating improved imaging methods for cell cultures.
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 plaque regions through adaptive threshold processes and watershed algorithms, generating Phase Gradient images without distortion.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If bright field optics are used to image cells, then the imaging system is simple and easy to operate, but the contrast between cells and surrounding media is very low making it difficult to detect cell objects and discriminate between live and lysed cells
Solution Approach 1:
The patent introduces phase field information as an intermediary representation that bridges the gap between simple bright field imaging and complex cell analysis. By extracting phase field data from bright field images through algorithms, the system creates an enhanced representation that reveals cell boundaries and structural details without requiring complex optical components, thus maintaining operational simplicity while improving detection accuracy.
Solution Approach 2:
The patent replaces complex mechanical optical systems (such as phase contrast or dark field microscopy) with computational methods that process bright field images. Instead of using additional optical components to enhance contrast, the system uses image processing algorithms to extract phase field information, substituting mechanical optical complexity with computational intelligence.
2Measurement precision
If traditional imaging methods are used for plaque detection, then the process is laborious and requires stringent aseptic conditions, but the accuracy in distinguishing between live and lysed cells is insufficient
Solution Approach 1:
The system enables self-service monitoring of cell cultures through automated image analysis. The phase field extraction and processing algorithms automatically distinguish between live and lysed cells, detect plaques, and provide quantitative data without requiring manual intervention or stringent aseptic handling during imaging, thus improving both accuracy and productivity.
Solution Approach 2:
The patent implements feedback mechanisms where the image analysis system continuously monitors cell culture conditions, provides quantitative data on cell viability and plaque formation, and enables real-time detection of changes in cell morphology and density, allowing for adaptive experimental design and reduced manual monitoring requirements.
3Measurement precision
If phase contrast optics are used to improve cell visibility, then cell contrast and structural details are enhanced, but the system complexity increases and requires more sophisticated optical components
Solution Approach 1:
The patent creates a computational copy or representation of the phase field information from bright field images. Instead of using complex optical systems to directly capture phase information, the system creates a digital replica of the phase field through image processing algorithms, achieving the same analytical capability with simpler hardware.
Solution Approach 2:
The patent changes the parameter representation by extracting phase field information as a separate computational parameter from the intensity information in bright field images. This allows the system to analyze cell structure and boundaries using phase field data derived from simple bright field imaging, avoiding the need for complex phase contrast optics while maintaining structural detail resolution.
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 method provides objective data for cell culture monitoring, ensures consistent treatment of cell lines, and enables accurate confluence measurements, cell morphology analysis, and quantitative assessment of cell growth, facilitating improved decision-making and process repeatability across labs.
Implementation Method 1
Phase contrast optics utilizes the different phase delay of the inner material and the surrounding media
Implementation Method 2
This behavior is the phenomena behind the Transport of Intensity Equation methodology for recovering the phase of the bright field illuminated subjects
Implementation Method 3
Live cells with an organized shape concentrate the illumination, forming bright spots in the above focus regions of the field
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.