Flow Cytometer PMT Sensitivity Adjustment via Reference Control
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Solution Overview
Problem
In flow cytometers, adjusting the detection sensitivity of photomultiplier tubes (PMTs) for different fluorescent dyes requires establishing a functional relationship through numerous measurements, especially when new PMTs or dyes are introduced, leading to inefficiencies and increased measurement time.
Innovation Solution
A measuring apparatus and method that allows for easy adjustment of detection sensitivity by using a control unit to adjust the voltage applied to PMTs based on reference values and statistical analysis of fluorescence intensities from control samples, thereby reducing the need for extensive recalibration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the functional relationship between applied voltage and output fluorescence value is determined through numerous measurements for each new PMT or fluorescent dye, then the detection sensitivity adjustment precision is improved, but the time and effort required for recalibration increases
Solution Approach 1:
The patent applies preliminary action by pre-determining the functional relationship between applied voltage and output fluorescence value using a reference PMT and reference fluorescent dye before actual measurements. The control unit stores this预先 determined relationship and uses it to automatically calculate the required applied voltage for target fluorescence values, eliminating the need for numerous measurements when new PMTs or dyes are introduced.
Solution Approach 2:
The patent uses a reference PMT and reference fluorescent dye as standard copies to establish the functional relationship. This reference relationship is then applied to new PMTs and dyes through automatic calculation by the control unit, avoiding the need to recreate the functional relationship through extensive measurements for each new component.
2Measurement precision
If the detection sensitivity is adjusted by experimentally determining the functional relationship through many measurements, then the accuracy of fluorescence measurement is improved, but the complexity of the adjustment process increases
Solution Approach 1:
The control unit automatically performs the sensitivity adjustment process by calculating the required applied voltage based on the target fluorescence value and the pre-determined functional relationship. This self-service capability eliminates the need for manual experimental determination and reduces the complexity of the adjustment process while maintaining measurement accuracy.
Solution Approach 2:
The patent replaces the manual mechanical adjustment process with an automated calculation system. The control unit uses the pre-determined functional relationship to automatically calculate the optimal applied voltage, substituting the complex manual experimental process with a simple computational approach.
3Adaptability or versatility
If numerous measurements are performed to obtain data for new PMT or fluorescent dye, then the adaptability to new components is improved, but the productivity during setup decreases
Solution Approach 1:
The patent creates a universal functional relationship using a reference PMT and reference fluorescent dye that can be applied to various new PMTs and fluorescent dyes. This universal approach allows the system to adapt to new components without requiring numerous new measurements, thereby maintaining high setup productivity while ensuring broad adaptability.
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 approach enables rapid and precise adjustment of detection sensitivity, reducing the time and effort required for recalibration when new PMTs or fluorescent dyes are used, while maintaining accurate fluorescence measurements.
Implementation Method 1
The present invention relates to a measuring apparatus... for irradiating a laser light on a fluorescence-labeled sample to measure the fluorescence generated from each cell... uses a fluorescence detector such as a photomultiplier tube (PMT)... capable of amplifying the detected signal
Implementation Method 2
The detection sensitivity of each fluorescence detector, that is, the degree of signal amplification, is adjusted according to the fluorescent dye to be detected
Data Source
Figure 1A~1B
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AI summary
PROBLEM: To easily adjust a measuring apparatus for detecting fluorescence from a sample. SOLUTION: A measuring apparatus 100 includes a flow cell 10 through which a sample containing particles flows, a light source 20 for irradiating light on the sample flowing through the flow cell 10, a fluorescence detector 30 for detecting the fluorescence generated from the sample irradiated with light from the light source 20, and a control unit 40 for flowing a positive control sample containing a fluorescent dye through the flow cell 10, measuring the fluorescence generated from the positive control sample irradiated by the light from the light source 20 via the fluorescence detector 30, comparing the obtained measurement value and a reference value, and adjusting the detection sensitivity of the fluorescence detector 30 according to the comparison result.