Bacteria Detection via Fluorescent Staining and Settling
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Solution Overview
Problem
Current methods for identifying and quantifying bacteria in samples face challenges such as difficulty in differentiation from debris, lack of standardization, and limited precision, especially when using glass slide interpretations.
Innovation Solution
A method involving staining a liquid sample with a fluorescent stain, allowing components to settle, and then imaging a Z-axis field of view using a confocal or epifluorescence microscope to determine the presence, type, and quantity of bacteria, while correlating these findings to the entire sample.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Difficulty of detecting and measuring
If traditional brightfield microscopy is used to visualize bacteria in bulk fluid, then bacteria can be detected, but they cannot be differentiated from debris and dust due to similar size and minimal morphologic variation
Solution Approach 1:
The patent applies fluorescent staining to bacteria, causing them to emit light at specific wavelengths that distinguish them from non-fluorescent debris and dust. This color/fluorescence change enables clear differentiation and accurate identification of bacteria in the sample.
Solution Approach 2:
The patent introduces a fluorescent stain as an intermediary substance that selectively binds to bacteria. This intermediary enhances the detectability and differentiability of bacteria from background debris, solving the identification accuracy problem.
2Quantity of substance
If glass slide interpretation methods are used for bacteria quantification, then bacteria can be counted, but the methods lack standardization and precision
Solution Approach 1:
The patent replaces manual glass slide interpretation with automated flow cytometry. This substitution of mechanical/manual analysis with automated optical and electronic measurement systems provides standardized, precise, and reproducible bacteria quantification.
Solution Approach 2:
The patent employs automated flow cytometric analysis that objectively measures and counts bacteria based on their fluorescent properties. This self-service automated system eliminates inter-observer variability and provides consistent, precise quantification results.
3Measurement precision
If fluorescent staining and flow cytometry are used to improve bacteria detection accuracy, then identification precision improves, but the method complexity increases
Solution Approach 1:
The patent uses a multi-functional flow cytometer that can perform staining, sorting, counting, and characterization of bacteria in a single instrument. This multi-functionality reduces overall system complexity compared to using separate specialized instruments for each function.
Solution Approach 2:
The patent performs fluorescent staining of bacteria before flow cytometric analysis. This preliminary action prepares the sample in advance, allowing the complex flow cytometry instrument to focus only on detection and measurement, thereby managing overall method complexity.
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 method provides a simple, accurate, and quick means to identify and quantify bacteria, overcoming the limitations of traditional methods by enhancing precision and standardization.
Implementation Method 1
staining a liquid sample with a fluorescent stain for bacteria
Implementation Method 2
allowing the components in the liquid sample to settle for a predetermined amount of time
Data Source
AI summary
An apparatus and method for detecting the type, presence or amount of bacteria in a sample. The apparatus and method involve staining a liquid sample with a fluorescent stain corresponding to bacteria and imaging one or more portions of the sample at a selected set of sample Z-axis heights, X-Y coordinates, or both, within the sample after a predetermined time to allow for non-bacterial components in the sample to settle out of solution. A calibration method for correlating the concentration or amount of bacteria in biological samples by different analysis methods including glass slide methods, agar plate colony culture methods and high precision methods.