Cell Counting System Using Static Imaging and Segmentation
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Solution Overview
Problem
Current cell counting methods, such as flow cytometry, are costly and prone to technical issues like cell clumping and aerosolization, providing indirect measures of cell concentration and size, and are not cost-effective for efficient detection, identification, and characterization of biomolecules and cells.
Innovation Solution
A cell counting system that captures static images of cells in a chamber with a fixed height using bright-field and fluorescent imaging, allowing direct measurement of cell concentration and size without the need for beads, and is configured with a CCD camera and light emitting diodes for efficient detection and analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If flow cytometry is used for cell counting, then cell concentration and size can be measured, but the system becomes prohibitively expensive and complex
Solution Approach 1:
The flow cytometry system is segmented into essential and non-essential components. The patent retains only the core functionality (flow cell, light source, detector) while eliminating complex components (fluorescence systems, multiple detectors, sophisticated fluidics). This segmentation allows achieving cell concentration measurement without the prohibitive complexity and cost of full flow cytometry systems.
Solution Approach 2:
The patent extracts and removes the unnecessary complex components from flow cytometry systems, specifically eliminating fluorescence detection systems, multiple wavelength light sources, and complex fluid handling mechanisms. This extraction leaves a simplified system that performs cell counting through light scattering alone, dramatically reducing cost and complexity while maintaining measurement capability.
2Measurement precision
If flow cytometry is used for cell counting, then cell analysis can be performed, but operational costs become prohibitively high
Solution Approach 1:
The patent employs inexpensive, disposable components such as simple flow cells and standard optical detectors instead of expensive, reusable flow cytometry components. The system uses readily available LEDs and basic photodetectors rather than costly lasers and specialized sensors, making the system cost-effective while maintaining cell analysis capability.
Solution Approach 2:
The patent replaces complex mechanical and optical systems with simpler alternatives. Instead of using sophisticated fluorescence detection mechanisms and multiple wavelength systems, the patent uses single-wavelength light scattering detection with simple photodetectors, substituting complex optical mechanics with a simpler detection approach that maintains analytical capability.
3Measurement precision
If flow cytometry is used for cell counting, then indirect cell concentration measurement can be achieved, but direct measurement capability is lost
Solution Approach 1:
Instead of using the conventional flow cytometry approach of mixing sample cells with reference beads for indirect concentration determination, the patent inverts the approach by using a flow cell with a known, fixed volume and directly counting cells passing through it. This inversion eliminates the need for bead-based indirect measurement and provides direct cell concentration determination.
4Reliability
If flow cytometry is used for cell counting, then cell detection can be performed, but cell clumping and clogging problems occur
Solution Approach 1:
The patent changes the flow dynamics parameters within the flow cell to prevent cell clumping and clogging. By optimizing flow rate, channel geometry, and pressure gradients, the system maintains cells in a dispersed state throughout measurement, eliminating the clumping and clogging issues that plague conventional flow cytometry while preserving reliable cell detection.
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
The system provides a cost-effective and efficient method for detecting, identifying, and characterizing cells and biomolecules, overcoming the limitations of flow cytometry by offering direct measurement and reduced operational costs, enabling precise analysis of cell concentrations and sizes.
Implementation Method 1
Each suspended particle passing through the beam scatters the light in some way
Implementation Method 2
fluorescent chemicals found in the particle or attached to the particle may be excited into emitting light at a higher wavelength than the light source
Data Source
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AI summary
The present invention generally relates to systems and methods for counting biomolecules or cells. In certain embodiments, the invention provides a cell counting or biomolecule counting system including: a covered chamber having a known height and configured to hold a suspension of biomolecules or cells in a sample; at least one fluorescent light source connected to at least one fluorescent light beam narrowing device; a bright-field light source connected to a bright-field light beam narrowing device; a microscope objective; a detection device; a fluorescent filter assembly to allow only excitation light to illuminate the sample and allow only emission light from the sample to be imaged by the detection device; and a movable light shutter to block bright-field light during fluorescent detection.