Endotoxin Quantification Using Real-Time Luminescence Calibration
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current endotoxin detection methods in biological samples are time-consuming, require manual handling steps leading to variable results, and rely on fixed measurement periods that may be longer than necessary or too short, with errors only detected at the end of the process.
Innovation Solution
A method using an analytical instrument with an imager and analytical support that monitors the temporal evolution of endotoxin concentration through luminescence reactions in biological and reference chambers, determining calibration relationships at each measurement instant to calculate endotoxin concentrations continuously during the measurement period.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If a fixed measurement period is used for all measurements, then the measurement process is simplified and standardized, but the measurement time is longer than necessary for most measurements and may still be too short for some specific cases
Solution Approach 1:
The patent implements dynamic measurement time adaptation by monitoring the temporal evolution of luminescence signals in real-time. The measurement process automatically adjusts its duration based on when the signal reaches a stable plateau, allowing each measurement to use the minimum necessary time rather than a fixed predetermined period. This resolves the contradiction by making the measurement time dynamic rather than static.
Solution Approach 2:
The system continuously monitors the luminescence signal evolution and uses this feedback to determine when to stop the measurement. When the signal reaches a stable plateau indicating complete reaction, the measurement automatically terminates. This feedback mechanism allows the system to adapt measurement duration to actual reaction kinetics, eliminating unnecessary waiting time while ensuring complete reactions.
2Adaptability or versatility
If manual preparation steps are used for standard dilutions and internal controls, then flexibility in test design is maintained, but the process becomes time-consuming and results become variable
Solution Approach 1:
The system performs automated preparation of standard dilutions and internal controls using robotic liquid handling. The automated system prepares all necessary dilutions and controls without manual intervention, eliminating the time-consuming and variable manual preparation steps while maintaining the ability to design different test configurations through software programming.
Solution Approach 2:
The patent replaces manual mechanical preparation operations with automated robotic liquid handling systems. This substitution eliminates human variability in preparation steps while maintaining flexibility through programmable test designs, resolving the contradiction between manual flexibility and automated precision.
3Measurement precision
If measurement errors are detected only at the end of the fixed measurement period, then the complete reaction data is available for analysis, but errors cannot be corrected and time is wasted on invalid measurements
Solution Approach 1:
The system implements continuous real-time monitoring of luminescence signals throughout the measurement period. Quality control parameters are evaluated continuously, and when errors or anomalies are detected, the system immediately flags the measurement as invalid and can terminate early. This continuous feedback allows error detection at the moment it occurs rather than waiting for the end of the fixed measurement period.
Solution Approach 2:
The system performs preliminary quality checks and validation during the measurement process itself, before the measurement period concludes. By monitoring signal evolution in real-time and applying quality control criteria continuously, the system can identify and correct issues early, preventing waste of time on completely invalid measurements.
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 reliable, real-time monitoring of endotoxin concentration, allowing for early detection of measurement errors, reducing analysis time, and ensuring accurate results by stopping the process when stability is achieved.
Implementation Method 1
the reference chambers being provided with reference reagents and different concentrations of endotoxins, said reference reagents being capable of causing a luminescence reaction in the presence of the reference liquid depending on the concentration of endotoxins
Implementation Method 2
the analysis chamber being equipped with analytical reagents capable of causing a luminescence reaction in the presence of endotoxins in the biological sample
Data Source
Figure 1
Figure 2
Figure 3
AI summary
A method for quantifying endotoxins in a biological sample with an analysis medium added to the field of view of an imager, the analysis medium comprising at least one analysis chamber and a plurality of reference chambers, comprising the steps of, for each of a plurality of measuring times of a measuring period: a) acquiring (S02) an image and determining (S03) light intensity valves in the reference and analysis chambers; b) determining (S04) a calibration relationship connecting the light intensity value and the concentration at this measuring time; c) determination (S05) of at least one endotoxin concentration in a biological sample from said calibration relationship and the intensity value in the analysis chamber at said measurement time; the method subsequently comprising the step of d) determining (S06) a temporal evolution of an endotoxin concentration in the biological sample over the course of the measuring period from the endotoxin concentration for a plurality of measurement times.