Viability Staining Method Using Fluorescent Dye and Quencher
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for detecting microbial contamination in industries like food, water, and pharmaceuticals are slow and lack specificity, requiring days to determine the presence and viability of microbial cells, which can lead to product quarantines and increased costs.
Innovation Solution
A method using a membrane permeable nucleic acid binding fluorescent dye and a membrane impermeable quencher to selectively label and differentiate viable and non-viable cells, allowing for rapid detection of viable microorganisms through fluorescence emission analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional cell culture methods are used to assess microbial contamination, then the detection is thorough and accurate, but the testing time is extended to several days
Solution Approach 1:
The patent replaces traditional mechanical cell culture methods with a fluorescence-based optical detection system. The method uses fluorescent dyes that bind to nucleic acids in viable cells, allowing rapid visualization and counting under a fluorescence microscope within hours rather than days, while maintaining detection accuracy.
Solution Approach 2:
The patent employs fluorescent dyes that emit specific wavelengths of light when bound to nucleic acids in viable cells. This color/fluorescence change enables rapid differentiation between viable and non-viable cells through fluorescence microscopy, providing quick results without lengthy culture periods.
2Measurement precision
If nucleic acid binding stains are used for mammalian cells, then the staining works well for mammalian cells, but the staining effectiveness decreases for microbial cells due to lower nucleic acid content
Solution Approach 1:
The patent modifies the staining parameters by using fluorescent dyes with high affinity for nucleic acids and optimizing the staining protocol to work with the lower nucleic acid content in microbial cells. The method adjusts dye concentration and exposure time to ensure effective staining of both mammalian and microbial cells despite their different nucleic acid quantities.
Solution Approach 2:
The patent develops a universal staining method that can effectively stain both mammalian cells and microbial cells using the same fluorescent nucleic acid binding dyes. The protocol is designed to be adaptable across different cell types by adjusting parameters such as dye concentration and incubation time, making it versatile for various applications.
3Productivity
If rapid detection methods are implemented, then the quarantine time is reduced and productivity increases, but the detection precision and reliability may be compromised
Solution Approach 1:
The patent replaces time-consuming mechanical culture methods with rapid fluorescence-based optical detection. The fluorescent dyes provide immediate visualization of viable cells, enabling results within hours while maintaining reliability through the specific binding of dyes to nucleic acids in metabolically active cells.
Solution Approach 2:
The patent uses fluorescent dyes as intermediaries that specifically bind to nucleic acids in viable cells. This intermediary approach allows rapid detection by providing a visible signal through fluorescence microscopy, enabling quick results without compromising reliability due to the specific and selective nature of the dye-nucleic acid interaction.
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 of viable microbial cells within hours, reducing quarantine times and improving the efficiency of microbial contamination testing, while maintaining the viability of cells for further identification.
Implementation Method 1
a membrane permeable nucleic acid binding fluorescent dye under conditions that permit the fluorescent dye to permeate both viable and non-viable cells
Implementation Method 2
a membrane impermeable nucleic acid binding fluorescence quencher capable of quenching fluorescence produced by the fluorescent dye
Data Source
Figure 1A
Figure 1B
Figure 1C
AI summary
The invention relates to a method of detecting viable cells in a cell sample, using a membrane permeable fluorescent label that permeates both viable and non- viable cells and a membrane impermeant quencher that selectively permeates non-viable cells.