Slow-Acting Decolorizing Formulation for Automated Gram Staining
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Solution Overview
Problem
Automated Gram staining methods face inconsistencies due to the rapid decolorizing step, which is too fast for automated systems, leading to inadequate decolorization of Gram-negative bacteria and over-decolorization of Gram-positive bacteria.
Innovation Solution
The use of slow-acting decolorizing formulations, such as those containing 1,2-propanediol or ethylene glycol, which prolong the decolorizing process, allowing for selective removal of the primary stain-trapping agent complex from Gram-negative bacteria without affecting Gram-positive bacteria, enabling automation of the Gram staining process.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If a fast-acting decolorizing agent (alcohol or acetone) is used, then decolorization of Gram-negative bacteria is achieved quickly, but the process is too fast for automated staining systems and leads to inconsistent results
Solution Approach 1:
The patent changes the chemical parameters of the decolorizing agent by using slower-acting formulations (reducing alcohol/acetone concentration) and adjusts the duration parameter to extend exposure time from seconds to minutes, enabling automated systems to achieve consistent decolorization of Gram-negative bacteria without over-decolorizing Gram-positive bacteria
2Extent of automation
If the decolorizing exposure time is extended for automated processing, then automation is enabled, but over-decolorization of Gram-positive bacteria occurs
Solution Approach 1:
The patent modifies the decolorizing formulation parameters (using slower-acting agents with reduced alcohol/acetone concentration) to match the extended time parameters required by automated systems, thereby maintaining staining accuracy while enabling automation
Solution Approach 2:
The patent introduces a specially formulated decolorizing agent that acts as an intermediary between the rapid manual process and the slow automated process, providing controlled decolorization kinetics that prevent over-decolorization while enabling automation
3Measurement precision
If manual Gram staining is performed, then the decolorizing step can be controlled precisely, but the process cannot be automated
Solution Approach 1:
The patent designs a decolorizing formulation that self-regulates the decolorization process through its slower kinetics, eliminating the need for manual timing control and enabling automated systems to achieve precision comparable to manual methods
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 approach ensures consistent and accurate differentiation between Gram-positive and Gram-negative bacteria, maintaining the staining of Gram-positive bacteria while effectively decolorizing Gram-negative bacteria, thus enabling reliable automated Gram staining.
Implementation Method 1
contacting the sample with a slow-acting decolorizing formulation (such as one that includes 1,2-propanediol or ethylene glycol) under conditions sufficient to significantly remove a primary stain-trapping agent complex from Gram-negative bacteria
Data Source
AI summary
Provided herein are methods of staining biological material for the purpose of detecting, and in some examples also identifying, microorganisms. Methods of Gram staining bacteria using a slow-acting decolorizing formulation, such as one that includes 1,2-propandiol or ethylene glycol, can be used to extend the time of the decolorizing step, and thus permit automation of the Gram staining method. Also provided are compositions and kits for performing automated Gram staining on microscope slides.