Stabilized Hematoxylin Composition for Automated Staining

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Solution Overview

Problem

Hematoxylin staining solutions for biological samples face instability issues due to changes in hematein concentration over time, leading to staining inconsistencies, particularly in automated processes where visual adjustments are costly and disruptive.

Innovation Solution

A stabilized hematoxylin composition is developed, including a solvent, hematoxylin, a chemical oxidant to convert hematoxylin to hematein, a mordant, cyclodextrin, and hydroquinone, which enhances stability and maintains effective dye concentration for consistent staining.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Speed

If chemical oxidant is used to accelerate conversion of hematoxylin to hematein, then staining speed is improved, but solution stability deteriorates due to faster degradation

Engineering Contradiction:
Improveconversion speed of hematoxylin to hemateinVSAvoidsolution stability
Core Design Contradiction:
SpeedVSStability of the object's composition

Solution Approach 1:

The patent changes the chemical parameters of the solution by adding specific stabilizing agents (sulfite, bisulfite, or dithionite) that alter the oxidation-reduction balance. These agents donate electrons to hematein, preventing its over-oxidation to degraded products, thereby maintaining solution stability while allowing accelerated conversion to proceed

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The stabilizing agents (sulfite, bisulfite, or dithionite) act as intermediary substances that mediate between the chemical oxidant and hematein. They provide a controlled electron transfer mechanism that prevents direct over-oxidation of hematein, serving as a protective intermediary layer in the chemical reaction system

Inventive Principle:
Principle #24Intermediary (Mediator)

2Manufacturing precision

If visual inspection is used to compensate for hematein concentration changes, then staining consistency is improved, but device complexity and cost increase due to imaging equipment requirements

Engineering Contradiction:
Improvestaining consistencyVSAvoidimaging equipment complexity
Core Design Contradiction:
Manufacturing precisionVSDevice complexity

Solution Approach 1:

The hematoxylin solution becomes self-regulating through the inclusion of stabilizing agents that automatically maintain hematein concentration within an effective range. The solution self-corrects for concentration changes over time without requiring external monitoring or adjustment, making the system self-sufficient and eliminating the need for complex imaging equipment

Inventive Principle:
Principle #25Self-service

Solution Approach 2:

The patent extracts and eliminates the need for complex imaging and monitoring equipment by fundamentally stabilizing the chemical composition. Instead of adding complex detection systems to compensate for instability, the solution itself is made stable through chemical modification, removing the harmful dependency on expensive equipment

Inventive Principle:
Principle #2Taking out (Extraction)

3Ease of operation

If hematein concentration is allowed to change over time, then manual adjustment is possible, but productivity decreases due to extended processing time and sample reprocessing

Engineering Contradiction:
Improvemanual adjustment flexibilityVSAvoidstaining throughput
Core Design Contradiction:
Ease of operationVSProductivity

Solution Approach 1:

The stabilizing agents ensure continuous maintenance of effective hematein concentration throughout the solution's shelf life and during use. This continuous stability eliminates interruptions, reprocessing steps, and time losses associated with concentration variability, enabling continuous high-throughput processing in automated systems

Inventive Principle:
Principle #20Continuity of useful action

Solution Approach 2:

The stabilizing agents are incorporated into the solution during preparation, performing preliminary stabilization action before the solution is put into use. This advance preparation ensures that the solution maintains optimal staining characteristics from the first use through its entire lifespan, preventing future consistency issues

Inventive Principle:
Principle #10Preliminary action

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

The stabilized composition provides improved stability and darker staining of biological samples, especially in automated methods, reducing the need for costly imaging and minimizing processing disruptions by maintaining hematein concentration over time.

Implementation Method 1

an oxidation product of hematoxylin, hematein, becomes the active staining component

Methodology Applied
Scientific EffectOxidation: Oxidation

Implementation Method 2

both of cyclodextrin and hydroquinone

Methodology Applied
Scientific EffectHost-guest complexation: Adsorption

Implementation Method 3

both of cyclodextrin and hydroquinone

Methodology Applied
Scientific EffectReduction: Reduction

Data Source

PatentEP2130023B1Stabilized hematoxylin
Publication Date: 2015.05.06 VENTANA MEDICAL SYSTEMS INC
  • EP2130023B1 patent drawingFigure 1
  • EP2130023B1 patent drawingFigure 2
  • EP2130023B1 patent drawingFigure 3

AI summary

A stabilized hematoxylin composition is disclosed that includes one or both of a host compound and an antioxidant. The disclosed composition exhibits sufficient stability to be utilized in an automated staining process without undue degradation prior to use of the composition to stain a biological sample. Methods of using and making the stabilized composition also are disclosed.