Flow Cytometer Filter Profiling and Calibration
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Solution Overview
Problem
Flow cytometry systems face challenges in efficiently capturing and quantifying fluorescent light emitted by small particles due to the need for high-intensity stimulating light, omnidirectional emission of fluorescing light, and the faintness of the emitted light, which can be beyond the detection capabilities of conventional photodetector systems.
Innovation Solution
A flow cytometry system that includes a calibration light source emitting different spectral profiles, which are directed through focusing optics and optical filter elements to detectors, allowing for the determination of filtering characteristics and optimization of light capture and detection, using processors to analyze output data and adjust settings for improved measurement efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Illumination intensity
If high-intensity stimulating light is used to cause fluorescence emission at detectable levels, then the fluorescence signal intensity is improved, but the energy consumption increases significantly
Solution Approach 1:
The patent applies local quality by using hydrodynamic focusing to concentrate the stimulating light and fluorescent emission into a tightly focused sample stream. This localized concentration of light intensity at the sample position enables detectable fluorescence signals while minimizing the overall energy required from the light source, as the high intensity is only needed where the particles are located rather than throughout the entire illumination field.
2Device complexity
If conventional photodetector systems are used to detect fluorescent light, then the system complexity is kept simple, but the detection capability is insufficient for faint fluorescent signals
Solution Approach 1:
The patent employs photomultiplier tubes as intermediary devices that bridge the gap between faint fluorescent signals and detectable electrical outputs. These photodetectors convert the weak optical signals into electrical currents through the photoelectric effect, and subsequent electron multiplication amplifies these signals by multiple orders of magnitude, enabling detection of extremely faint fluorescent emissions without requiring complex alternative detection systems.
3Ease of operation
If optical filter elements are not precisely characterized, then the system operation is simple, but the measurement accuracy deteriorates due to misconfiguration
Solution Approach 1:
The patent implements preliminary action by performing comprehensive spectral characterization of optical filter elements before they are installed in the flow cytometer. The system measures the transmission spectrum of each filter using a spectrometer and stores these characteristics in a database. This pre-characterization ensures that when filters are later installed or replaced, the system can automatically retrieve and apply the correct spectral data, eliminating the need for manual re-characterization and preventing measurement errors due to misconfiguration.
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
Enhances the ability to accurately detect and quantify fluorescent light by optimizing the filtering characteristics of the system, improving measurement efficiency and reducing errors associated with misconfiguration of optical filters.
Implementation Method 1
each optical path passes through a corresponding one or more optical filter elements and terminates at a corresponding detector
Implementation Method 2
extremely sensitive photodetector systems may be used, such as photomultiplier tubes, which convert the received florescent light into an electrical current that may be amplified by multiple orders of magnitude
Implementation Method 3
a fluorescing indicator, that may be stimulated to provide a quantifiable response, e.g., to emit light that may be detected by optical sensors
Data Source
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AI summary
Disclosed is a system and method for characterizing optical filters in a flow cytometer and optionally checking the operation of detectors in the flow cytometer. In some embodiments, the system may utilize an LED board having an opening through which the fluorescence and side scatter beams, rays, or images pass and light emitting diodes around the opening that emit light having different spectral profiles. The different spectral profiles allow the system to identify the optical filters that are placed in the flow cytometer, to verify detector operation, to assist in instrumentation troubleshooting, and to provide a quantitative reference for detector comparison.