Rectangular Flow Cell Optics for High-Volume Flow Cytometry
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Solution Overview
Problem
Traditional flow cytometry systems are inadequate for detecting low concentrations of analyte entities in high volume fluid samples due to low flow rates and limited sensitivity, making them unsuitable for bioburden analysis in liquids such as environmental water samples.
Innovation Solution
A flow cytometry system with a rectangular flow cell and spherical reflector configuration, using a dye combination for broad analyte detection, and omitting Mie scattering analysis to enhance discrimination between target and non-target particles, allowing for high volume analysis at increased flow rates.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional flow cytometry uses small flow rates (about 100 μL/min) with sheath flow through an analytical flow cell, then the system can obtain data without extraneous signal problems from the periphery of the glass and flow cell walls, but the flow rate is too slow for analyzing low concentration samples in high volumes
Solution Approach 1:
The patent removes the sheath flow component from the traditional flow cytometry system. By eliminating the sheath flow requirement, the system can process samples at much higher flow rates (up to 100 mL/min or more) without the signal quality degradation that occurs in traditional systems at high flow rates. This extraction of the sheath flow dependency enables high-volume sample analysis while maintaining detection capability.
Solution Approach 2:
The patent introduces a three-dimensional imaging approach using multiple detection angles (front scatter, side scatter, and fluorescent signals collected at different angles) to replace the traditional two-dimensional flow-based detection. This dimensional expansion allows the system to maintain measurement precision at high flow rates by capturing spatial information that compensates for the loss of flow-based signal stabilization.
2Measurement precision
If traditional flow cytometry uses sheath flow to focus the sample in the middle of the flow cell, then the interrogation light can be directed to the middle of the flow cell without extraneous signal problems, but the system cannot detect low numbers of analyte entities in high flow rates or high volume interrogation rates
Solution Approach 1:
The patent extracts the sheath flow dependency from the system architecture. By removing the requirement for sheath flow to achieve proper sample positioning and signal quality, the system can operate at high volume interrogation rates (processing entire 100 mL samples in minutes rather than hours) while maintaining the ability to detect low numbers of analyte entities through its alternative focusing and detection methodology.
Solution Approach 2:
The patent replaces the mechanical sheath flow focusing system with an optical and computational approach. The system uses specific light interrogation geometries and image processing algorithms to achieve particle detection and discrimination without requiring mechanical fluid dynamic control via sheath flow, enabling high-speed processing while maintaining detection sensitivity.
3Reliability
If flow cytometry is used with low flow rates and low sensitivity, then the system works well for laboratory settings with prepared liquid samples, but the technology is not suitable for bioburden analysis of dilute concentrations in environmental water samples
Solution Approach 1:
The patent fundamentally changes the operational parameters of flow cytometry by eliminating sheath flow and implementing high-flow-rate operation combined with multi-angle light scattering detection. This parameter transformation enables the system to process entire 100 mL water samples in minutes, providing both the reliability needed for bioburden analysis (detection of single cells in dilute samples) and the productivity required for practical environmental monitoring applications.
Solution Approach 2:
The patent creates a universal flow cytometry system that can handle both traditionally prepared laboratory samples and raw environmental water samples without requiring pre-concentration or extensive sample preparation. The system's ability to process high volumes at high flow rates while maintaining detection sensitivity makes it applicable to diverse sample types and applications, from clinical diagnostics to environmental monitoring.
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 rapid detection of low concentrations of analytes in high volumes, achieving analysis times of a few hours instead of days, and improving discrimination between viable and non-viable cells or particles.
Implementation Method 1
The spherical reflector has a concave reflective surface that has a reflective direction that is positioned substantially orthogonal with the light path such that reflected light is reflected along a reflected path that is substantially orthogonal with the light path
Implementation Method 2
The flow cytometer can include at least one light absorbing member to inhibit reflections from passing through the exit aperture
Implementation Method 3
at least one light emitter configured to emit light in a light path
Data Source
AI summary
A flow cytometer can include: at least one light emitter configured to emit light in a light path; a rectangular flow cell having flow cell width that is substantially lateral to the light path and a flow cell depth that is longitudinal to the light path, wherein the light path has an interrogation width at the flow cell that is narrower than the flow cell width; and a spherical reflector positioned adjacent to the rectangular flow cell and having a concave reflective surface that has a reflective direction that is positioned substantially orthogonal with the light path such that reflected light is reflected along a reflected path that is substantially orthogonal with the light path. At least one light absorbing member is positioned at least partially around the reflected path to absorb reflected light at an angle to the reflected path.