H&E Stain Removal for Multi-Target Tissue Analysis
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for cancer diagnosis face challenges in analyzing multiple biological targets from limited sample sizes, particularly when using hematoxylin and eosin (H&E) stained slides, as the H&E stains interfere with subsequent immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) analyses, limiting the ability to perform detailed molecular assessments.
Innovation Solution
A method is developed to remove the fluorescent H&E signal from stained biological samples using a series of washes, charge transfer agents, hydrogen peroxide with metal salts, and photo-induced chemical bleaching (PICB) agents, allowing for the detection of multiple targets on the same tissue section without residual interference.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If H&E staining is performed on biological samples for morphological assessment, then diagnostic accuracy for cancer detection is improved, but the H&E stains interfere with subsequent IHC and FISH analyses
Solution Approach 1:
The patent removes H&E stains from tissue sections through a multi-step process involving differentiation in acid alcohol, washing in water, and treatment with oxidizing agents like hydrogen peroxide. This extraction of the harmful stain component allows the same tissue section to be used for subsequent IHC and FISH analyses without interference from the original H&E staining.
Solution Approach 2:
The patent discards the H&E stain by removing it through differentiation and washing steps, then recovers the tissue section for additional molecular analyses. This enables the same valuable tissue sample to be reused for multiple diagnostic purposes, maximizing the utility of limited biopsy materials.
2Quantity of substance
If multiple targets are analyzed using IHC or FISH on separate tissue sections, then molecular characterization is achieved, but limited sample size restricts the ability to perform various analyses on the same sample
Solution Approach 1:
The patent makes a single tissue section serve multiple functions by first performing H&E staining for morphological assessment, then removing the stain to enable subsequent IHC and FISH analyses. This multi-functional use of the same tissue section maximizes the diagnostic information obtained from limited biopsy samples.
Solution Approach 2:
The patent performs H&E staining and initial morphological assessment as a preliminary action before removing the stain for molecular analyses. This allows the tissue to be evaluated for both morphological and molecular characteristics in sequence, ensuring comprehensive diagnostic workup from a single sample.
3Adaptability or versatility
If H&E stains are removed from tissue sections to enable IHC and FISH, then molecular analysis feasibility is improved, but residual eosin fluorescence prevents IF and FISH detection
Solution Approach 1:
The patent uses strong oxidizing agents, particularly hydrogen peroxide, to treat the tissue sections after H&E staining removal. This oxidation process effectively eliminates residual eosin fluorescence that would otherwise interfere with fluorescence-based IHC and FISH detections, enabling clear fluorescence signal detection.
Solution Approach 2:
The patent converts the harmful residual eosin fluorescence into a benefit by using it as a target for oxidation. The hydrogen peroxide treatment specifically addresses the fluorescence interference problem, transforming the previously harmful residual stain into a non-interfering state that allows successful fluorescence-based molecular analyses.
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 enables the effective removal of H&E signals, enabling subsequent IHC and FISH analyses on the same tissue section, facilitating detailed molecular characterization and targeted therapy selection for cancer patients.
Implementation Method 1
photo-induced chemical bleaching (PICB) agents
Implementation Method 2
hydrogen peroxide with metal salts
Implementation Method 3
charge transfer agents
Data Source
Figure 1~2
Figure 3A~3B
Figure 4
AI summary
Methods comprising probing multiple targets in an H&E stained biological sample are provided. The methods include the steps of providing a hematoxylin and eosin stained biological sample containing multiple targets, optionally detecting H&E staining of the sample, removing the hematoxylin and eosin signals, and detecting additional features or targets in the biological sample. The detecting step may include performing the steps of binding at least one probe to one or more targets to the sample, detecting a signal from the probe and contacting the sample with a bleaching agent. The process of binding, detecting and bleaching may be iteratively repeated.