Static Cell Counting via Fluorescence and Bright-Field Imaging
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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 biohazardous samples.
Innovation Solution
A cell counting system that captures static images of cells in a chamber with a fixed height, using both bright-field and fluorescent light sources, allowing for direct measurement of cell concentration and size without the need for beads, and includes a movable light shutter for fluorescent detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If flow cytometry is used for cell counting, then cell concentration can be measured, but the system cost is prohibitively high ($150,000 to $500,000)
Solution Approach 1:
The patent creates a simplified copy of flow cytometry functionality using standard microscopy equipment. Instead of requiring expensive flow cytometers, the system uses a microscope with imaging capabilities to capture cell images in a counting chamber, achieving similar cell concentration measurement through image analysis algorithms that replicate flow cytometry's counting capability at a fraction of the cost
Solution Approach 2:
The patent replaces the mechanical fluid flow system of flow cytometry with a static imaging system. Instead of hydrodynamically focusing cells through a nozzle and using light scatter detection, the system uses optical microscopy with image capture and computational analysis to measure cell concentration, substituting mechanical flow dynamics with optical and computational methods
2Measurement precision
If flow cytometry is used for cell counting, then cell concentration can be measured, but technical problems result from cells clumping and clogging or sticking in the nozzle
Solution Approach 1:
The patent extracts the problematic fluid flow and nozzle components from the cell counting system. By using a static counting chamber where cells are imaged in place rather than flowing through a nozzle, the system eliminates the source of clumping and clogging issues while maintaining the ability to measure cell concentration through image-based counting methods
Solution Approach 2:
Instead of flowing cells through a system and detecting them in motion as in flow cytometry, the patent inverts the approach by placing cells in a static chamber and imaging them in place. This reversal of the measurement paradigm eliminates flow-related problems while achieving reliable cell concentration measurements through stationary image capture and analysis
3Measurement precision
If flow cytometry is used for cell counting, then cell concentration can be measured, but aerosolization of the sample prevents biohazardous samples from being sorted
Solution Approach 1:
The patent converts the potential harm of aerosolization into a benefit by using a closed imaging chamber system. The same sealed environment that prevents aerosol escape also maintains optimal imaging conditions, and the system can process biohazardous samples safely without requiring aerosol generation, turning a safety constraint into an operational advantage
Solution Approach 2:
The patent introduces a closed imaging chamber as an intermediary between the sample and the external environment. This chamber acts as a barrier that prevents biohazardous aerosols from escaping while allowing optical imaging to proceed, mediating between the need for safe sample handling and the need for accurate cell measurement
4Measurement precision
If flow cytometry is used for cell counting, then cell concentration can be measured, but only an indirect measure is provided requiring mixing with beads
Solution Approach 1:
The patent extracts the bead calibration step from the measurement process. By using a known volume chamber with direct imaging, the system eliminates the need for bead-based indirect measurement and calibration curves, obtaining cell concentration directly from image analysis of cells in a defined volume without requiring external reference particles
Solution Approach 2:
The patent creates a direct imaging copy of the cell population in a known volume, eliminating the need for indirect bead-based measurement. The system captures actual cell images in a chamber of known dimensions and calculates concentration directly from the image data, copying the physical reality of cell distribution without requiring intermediary bead standards
5Device complexity
If manual cell counting with hemacytometer is used, then cost is reduced, but productivity is low and manual enumeration is required
Solution Approach 1:
The patent implements self-service by enabling the system to automatically count and analyze cells without manual enumeration. The imaging system captures cell images and computational algorithms automatically identify, count, and calculate cell concentration from the images, making the system serve itself without requiring operator intervention for the counting process while maintaining low cost through the use of standard microscopy equipment
6Device complexity
If manual cell counting with hemacytometer is used, then cost is reduced, but measurement precision is limited by microscope variation
Solution Approach 1:
The patent creates digital copies of the cell population through high-resolution imaging, replacing the analog visual inspection process. The captured images serve as permanent records that can be analyzed with consistent computational algorithms, eliminating the variability introduced by different operators viewing through microscopes and achieving reproducible measurements across different users and sessions
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 system provides a cost-effective, efficient method for detecting, identifying, and quantifying cells, reducing errors and sample hazards, while enabling direct measurement of cell concentration and size, and can be used for various biological samples.
Implementation Method 1
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
Implementation Method 2
Each suspended particle passing through the beam scatters the light in some way
Data Source
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.