Fluorescent Membrane Microorganism Detection Method
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Solution Overview
Problem
Current methods for detecting and quantifying microorganisms in liquid samples are slow, often taking 12 to 36 hours, and require expensive equipment or techniques that can be lethal to the microorganisms, limiting their viability for further analysis or treatment.
Innovation Solution
A method involving a membrane to deposit microorganisms, allowing them to form microcolonies, followed by labeling with a fluorescent compound that binds to endogenous constituents, enabling rapid detection and quantification without affecting viability, using a simple optical device with magnification between 1 and 50 and optical resolution of 6-20 Mpix.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If gold standard methods (plating on solid culture medium) are used for detection, then measurement precision is improved, but loss of time increases to 12-36 hours
Solution Approach 1:
The patent applies preliminary action by pre-coating the detection membrane with fluorescent compounds and culture medium before sample application. This pre-preparation eliminates the need for time-consuming post-detection colony cultivation, enabling rapid detection within 1-3 hours while maintaining accuracy through the fluorescent labeling of endogenous constituents.
Solution Approach 2:
The patent replaces the mechanical/optical system of visual colony counting on solid media with a fluorescent labeling and detection system. The fluorescent compounds bind to endogenous constituents of microorganisms, allowing detection through fluorescence imaging rather than traditional colony visualization, thereby reducing detection time while preserving measurement precision.
2Measurement precision
If sophisticated and expensive equipment is used for detection, then measurement precision is improved, but device complexity increases
Solution Approach 1:
The patent employs a disposable detection membrane pre-coated with fluorescent compounds and culture medium, replacing expensive, complex, and reusable sophisticated detection equipment. This single-use membrane approach maintains high detection precision while eliminating the need for expensive instrumentation and reducing device complexity in the detection system.
Solution Approach 2:
The patent introduces fluorescent compounds as intermediaries that bind to endogenous constituents of microorganisms. These fluorescent labels serve as mediators between the microorganisms and the detection system, enabling accurate detection through simple fluorescence imaging rather than requiring complex analytical equipment.
3Measurement precision
If lethal detection techniques are used, then measurement precision is improved, but object-generated harmful factors increase due to loss of viability
Solution Approach 1:
The patent applies self-service by utilizing endogenous constituents naturally present within the microorganisms themselves as targets for fluorescent labeling. This approach detects microorganisms in their native state without requiring external fixation, staining, or lethal processing steps, thereby maintaining viability while achieving precise detection.
Solution Approach 2:
The patent replaces lethal chemical fixation and staining procedures with a non-invasive fluorescent labeling method that binds to endogenous constituents. This substitution eliminates harmful factors that would otherwise kill the microorganisms, allowing detection with preserved viability for subsequent culturing or experimentation.
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 detection and quantification of microorganisms in under 8 hours without harming them, allowing for subsequent antibiotic susceptibility testing and further growth, reducing the need for expensive equipment and shortening diagnostic and therapeutic delays.
Implementation Method 1
putting the lower surface of said membrane into contact with a fluorescent compound capable of labeling an endogenous constituent of the microorganisms
Data Source
Figure 1
Figure 2A~2B
AI summary
The present invention concerns a method for detecting and optionally quantifying microorganisms in a liquid sample, comprising the step of (a) passing the liquid sample through a membrane thereby depositing the potentially present microorganisms on the upper surface of said membrane, (b) putting the lower surface of said membrane into contact with a culture medium and incubating the whole under conditions suitable for promoting the formation of microcolonies from the microorganisms, (c) putting the lower surface of said membrane into contact with a fluorescent compound capable of labeling an endogenous constituent of the microorganisms in order to label potentially formed microcolonies, (d) washing the lower surface of said membrane thereby eliminating the excess of fluorescent compound, and (e) detecting and optionally quantifying the potentially formed microcolonies labeled with the fluorescent compound, wherein the detection is performed using an optical device having a magnification between 1 and 50 and an optical resolution comprised between 6-20 Mpix.