Eosin Counterstain Sequence for Consistent H&E Tissue Staining

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Solution Overview

Problem

Existing automated staining machines often produce inconsistent and variable results in tissue staining, particularly with hematoxylin and eosin (H&E) stains, affecting diagnostic accuracy and reproducibility.

Innovation Solution

The method involves altering the sequence and application of acid, alkaline, wash, and transfer solutions during the staining process to improve consistency and reproducibility, including applying acid solutions at specific stages and using transfer solutions to minimize stain removal, thereby optimizing the application of primary and counterstains.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If automated staining machines are used to improve productivity and consistency, then staining efficiency increases, but staining consistency and reproducibility deteriorate due to variable results particularly in H&E stains

Engineering Contradiction:

Improvestaining efficiencyVSAvoidstaining consistency

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent applies parameter changes by precisely controlling the timing, sequence, and duration of acid solution applications during the staining process. By modifying when acid solutions are applied (immediately after counterstain vs. between primary and counterstain) and controlling contact times, the patent achieves both high productivity through automation and improved staining consistency, resolving the contradiction between automated efficiency and staining reliability

Inventive Principle:

Principle #35Parameter changes

2Measurement precision

If acid solutions are applied between primary staining and counterstaining to improve stain differentiation, then diagnostic clarity improves, but counterstain intensity and consistency deteriorate

Engineering Contradiction:

Improvediagnostic clarityVSAvoidcounterstain consistency

Core Design Contradiction:

Measurement precisionVSReliability

Solution Approach 1:

The patent applies inversion by reversing the conventional sequence: instead of applying acid solution between primary and counterstain, the acid solution is applied immediately after counterstaining. This inverted approach maintains counterstain intensity and consistency while still achieving adequate differentiation, thereby resolving the contradiction between diagnostic clarity and counterstain reliability

Inventive Principle:

Principle #13The other way round (Inversion)

3Manufacturing precision

If multiple wash solutions are applied after counterstaining to improve stain removal, then excess stain is reduced, but staining time and process complexity increase

Engineering Contradiction:

Improvestain uniformityVSAvoidprocess complexity

Core Design Contradiction:

Manufacturing precisionVSDevice complexity

Solution Approach 1:

The patent applies extraction by removing unnecessary wash steps from the staining process. By applying acid solution immediately after counterstaining and eliminating intermediate wash steps, the patent achieves adequate excess stain removal while simplifying the overall process, thus resolving the contradiction between stain uniformity and process complexity

Inventive Principle:

Principle #2Taking out (Extraction)

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 enhances the consistency and reproducibility of tissue staining, improving diagnostic clarity and reducing variability in H&E staining outcomes.

Implementation Method 1

apply an acid solution to the specimen immediately following staining with the counterstain

Methodology Applied

Scientific EffectAcid solution treatment:

Implementation Method 2

contacting the specimen with an alkaline solution

Methodology Applied

Scientific EffectAlkaline solution treatment:

Implementation Method 3

contacting the specimen with a wash solution to at least partially remove the staining reagent from the specimen

Methodology Applied

Scientific EffectWashing:

Implementation Method 4

apply at least two aliquots of transfer solution between counterstaining and the application or an organic fluid solution or a coverslip

Methodology Applied

Scientific EffectNeutral transfer:

Data Source

PatentUS20250277723A1Eosin staining techniques

Publication Date: 2025.09.04 VENTANA MEDICAL SYSTEMS INC

AI summary

Presented herein are methods of improving the consistency of staining with a counterstain. In some embodiments, the method makes use of an automated specimen processing apparatus or other staining device. In some embodiments, the staining methods are applied manually. In some embodiments, the counterstain includes cosin.