2D Fluorescence Spectroscopy in Agitated Microtiter Plates
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Solution Overview
Problem
Existing methods for measuring process parameters in microbial cultures using 2D fluorescence spectroscopy are limited to substances with fluorescence activity and cannot determine physiological states or substance transfer rates efficiently, requiring lengthy measurement times.
Innovation Solution
A method and device employing 2D fluorescence spectroscopy in agitated microtiter plates, where monochromatic excitation light is modified step-by-step, and emission spectra are recorded using a sensor matrix, allowing for the measurement of process parameters in non-fluorescent substances and physiological states, with a significant reduction in measurement time through a beam guidance system and optical coupler configuration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If conventional fluorescence measurement methods are used, then measurement of fluorescent substances is possible, but measurement of non-fluorescent substances and determination of physiological states cannot be performed
Solution Approach 1:
The patent transitions from conventional single-wavelength fluorescence measurement to 2D fluorescence spectroscopy, adding the excitation wavelength dimension to the measurement. This enables detection of both fluorescent and non-fluorescent substances by recording fluorescence emission across multiple excitation wavelengths, thereby expanding measurement versatility and recovering physiological state information that was previously inaccessible.
Solution Approach 2:
The invention systematically varies the excitation wavelength parameter across a range of values while measuring fluorescence emission. This parameter change enables the detection of substances with different fluorescence characteristics, including non-fluorescent substances that may exhibit fluorescence under specific excitation conditions, thus improving adaptability without losing physiological state information.
2Productivity
If conventional measurement methods are used, then simple equipment operation is maintained, but measurement time is excessively long
Solution Approach 1:
The patent employs a pre-calibrated lookup table that stores fluorescence spectra characteristics obtained from prior measurements. During actual measurement, the system quickly compares acquired spectra against this pre-prepared reference data, enabling rapid identification and quantification of substances without performing full spectral analysis in real-time, thus dramatically reducing measurement time while maintaining accuracy.
Solution Approach 2:
The invention creates a digital copy of fluorescence spectral characteristics in the form of a lookup table during a calibration phase. This copied reference data is then used for rapid comparison and identification during subsequent measurements, eliminating the need for time-consuming real-time spectral decomposition and significantly improving measurement speed.
3Adaptability or versatility
If 2D fluorescence spectroscopy is implemented, then comprehensive process parameter measurement is achieved, but device complexity increases
Solution Approach 1:
The patent introduces a lookup table as an intermediary data structure that mediates between the complex 2D fluorescence spectroscopy measurements and the final substance identification. This intermediary stores pre-computed spectral characteristics and enables rapid pattern matching, simplifying the interpretation process while maintaining the comprehensive detection capability of 2D spectroscopy.
Solution Approach 2:
The invention creates simplified digital representations (copies) of complex fluorescence spectral patterns in the lookup table. These copied spectral fingerprints allow the system to identify substances based on pattern matching rather than complex real-time spectral analysis, reducing computational complexity while preserving the ability to detect diverse substances including non-fluorescent ones.
4Quantity of substance
If parallel cultivation monitoring is performed, then data density is increased, but measurement time for each sample increases
Solution Approach 1:
The patent performs preliminary calibration and creates the lookup table once, storing all necessary spectral reference data. During parallel cultivation monitoring, the system only needs to perform quick spectral acquisitions and comparisons against the pre-prepared lookup table, enabling high-throughput measurement of multiple samples without proportionally increasing the time investment for each sample analysis.
Solution Approach 2:
The invention uses copied spectral reference patterns from the lookup table to rapidly identify and characterize substances in multiple parallel cultivations. This copying approach allows simultaneous or sequential measurement of many samples with minimal additional time per sample, as the complex spectral analysis has already been performed and stored during the initial calibration phase.
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 recording of 2D fluorescence spectra across multiple microreactors, reducing measurement time from hours to minutes, allowing for real-time monitoring of parallel cultivations and increased data density, and enabling the determination of respiratory activity without complex equipment.
Implementation Method 1
an automatically tunable monochromator (15) for spectral isolation of different wavelengths from the incident light of the light source (8)
Implementation Method 2
an optical element that decomposes the emission spectrum for each excitation wavelength into the different wavelengths and depicts the emission spectrum fanned out on a sensor matrix
Implementation Method 3
a photosensor is arranged on the other strand of the y-shaped optical waveguide
Implementation Method 4
an orbital shaker configured to agitate the liquid cultures by moving the at least one microtiter plate in an agitating motion
Data Source
AI summary
A method for measuring process parameters in liquid cultures in a plurality of microreactors of at least one microtiter plate includes continuously agitating the liquid cultures using an orbital agitator at least until the reaction is completed in all the microreactors. In order to allow process parameters also of such substances which themselves do not have any fluorescence activity to be measured with relatively low complexity and within a short time, 2D fluorescence spectra are recorded in a plurality of in particular different liquid cultures in the microreactors of agitated microplates. A device for carrying out the method is also disclosed.