Flow Cytometer Photodetector Calibration Light Source
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Solution Overview
Problem
Current methods for characterizing and calibrating photodetectors in flow cytometers, such as those using bead-based quality control and LED light sources, face limitations in precision, dynamics, and scalability, particularly in achieving optimal signal-to-noise ratio and dynamic detection range due to external factors and the need for complex feedback loops.
Innovation Solution
A device with separate control light sources connected in series to emission light sources, using identical LEDs for precise feedback regulation, allows for high-precision control of light pulses with desired signal shapes, enabling scalable and compact designs for multiple colors and intensity levels, thereby simplifying the characterization, standardization, and calibration of photodetectors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If bead-based quality control methods are used for PMT voltage optimization, then quality control can be performed, but the results are influenced by external factors (laser alignment, background noise) and intrinsic bead properties, preventing reliable optimization for actual experimental requirements
Solution Approach 1:
The patent uses a light source that replicates the actual fluorescence signal characteristics without requiring physical beads. This copying approach eliminates the influence of bead properties (size, fluorescence intensity variations) and external factors (laser alignment, background noise) that contaminate measurements when using actual beads for calibration.
2Adaptability or versatility
If the dynamic range of a PMT is increased to represent all events of interest, then more events can be detected, but the signal-to-noise ratio decreases, requiring PMT voltage reduction which lowers detection sensitivity
Solution Approach 1:
The patent employs a light source with dynamically adjustable intensity that can precisely control the output signal level. This allows systematic determination of the relationship between PMT voltage and both dynamic range and signal-to-noise ratio, enabling optimization of the operating point without the compromises required by static calibration methods.
3Manufacturing precision
If separate feedback control is implemented for each emission light source to ensure precise signal shape, then temporal precision is improved, but device complexity increases due to multiple feedback loops
Solution Approach 1:
The patent combines multiple emission light sources (with different spectral characteristics) into a single integrated light source assembly that couples to one optical fiber. This merging approach allows multiple colors to be emitted simultaneously or sequentially through a single coupling interface, reducing the number of separate feedback loops needed while maintaining precise temporal control through the shared optical path.
4Adaptability or versatility
If multiple emission light sources with different coupling efficiencies are used to provide different intensity levels, then dynamic range is improved, but the device size and complexity increase
Solution Approach 1:
The patent positions multiple emission light sources at different locations relative to the optical fiber coupling point, creating local variations in coupling efficiency. By strategically placing LEDs at different distances and angles from the fiber end, each light source naturally couples with different efficiency, providing multiple intensity levels without requiring additional optical components or increasing overall device size.
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 solution provides enhanced precision and flexibility in generating light pulses, allowing for comprehensive characterization and calibration of photodetectors, improving the signal-to-noise ratio and dynamic detection range, and enabling efficient calibration of photomultipliers in flow cytometers and microscopes.
Implementation Method 1
The device comprises light-emitting diodes which are excited to emit light pulses with a predetermined signal waveform
Implementation Method 2
separate, series-connected control light sources, the emission of which is detected by a feedback detector
Data Source
Figure 1~2B
Figure 3A~3B
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AI summary
The invention relates to a device for generating light pulses for characterizing, standardizing, and/or calibrating photodetectors, preferably within a flow cytometer or microscope. For this purpose, the device comprises emission light sources which are excited to emit light pulses with a predetermined signal waveform. The device is characterized by a feedback mechanism based on the provision of separate, series-connected control light sources, the emission of which is detected by a feedback detector. In preferred embodiments, the device comprises one or more emission groups with circularly arranged, multicolored emission light sources. To provide different intensity levels, the emission light sources or emission groups with different efficiencies can be coupled into an optical fiber.In further aspects, the invention also relates to uses of the device as well as systems or kits comprising the device.