Tissue Sample Staining for Simulated H&E and Multiplexed Imaging
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Solution Overview
Problem
Conventional pathology assessment methods using hematoxylin and eosin (H&E) staining interfere with subsequent multiplexed fluorescence imaging and are difficult to remove, making it challenging to analyze tissue samples effectively.
Innovation Solution
Simulated H&E imaging using curcumin and/or carmine to mimic hematoxylin and eosin staining, allowing for multiplexed fluorescence imaging without interference.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional H&E staining is applied to tissue samples, then morphology and structural information are provided for pathology assessment, but the stains interfere with subsequent multiplexed fluorescence imaging and are difficult to remove
Solution Approach 1:
The patent uses fluorescent analogs (curcumin for hematoxylin, eosin for eosin) that replicate the staining patterns and morphological information of conventional H&E stains, but emit fluorescence signals that can be detected without interference. This copying approach allows the same diagnostic information to be obtained while enabling subsequent multiplexed fluorescence imaging.
Solution Approach 2:
The patent changes the detection parameter from absorption-based optical microscopy (conventional H&E) to fluorescence-based detection. By using fluorescently labeled analogs that emit at specific wavelengths, the system maintains the morphological staining function while adding the capability for multiplexed fluorescence imaging without signal interference.
2Measurement precision
If conventional H&E staining is used, then tissue morphology is visualized, but sample integrity is compromised for subsequent fluorescence imaging steps
Solution Approach 1:
The fluorescent analogs replicate the morphological staining function of H&E while maintaining sample integrity. The fluorescent labels are designed to bind to the same tissue components as conventional H&E stains, providing identical morphological information without compromising the sample for subsequent fluorescence imaging or other molecular analyses.
Solution Approach 2:
The fluorescent analogs serve as intermediaries that bridge conventional H&E staining and fluorescence imaging. These analogs have dual functionality: they provide the morphological contrast needed for pathology assessment while simultaneously serving as fluorescence reporters that do not interfere with subsequent imaging steps.
3Loss of information
If multiple tissue sections are stained with different reagents, then comprehensive pathology information is obtained, but processing time and complexity increase
Solution Approach 1:
The patent merges the functions of multiple separate staining steps into a single multiplexed fluorescence staining procedure. By using fluorescent analogs that can be detected simultaneously at different wavelengths, the system obtains comprehensive pathology information (morphology from H&E analogs plus specific molecular markers) in one processing run rather than requiring sequential staining of multiple sections.
Solution Approach 2:
The fluorescent staining system provides multi-functionality by simultaneously delivering morphological information (through curcumin and eosin analogs) and specific molecular marker detection (through additional fluorescent probes). This universal approach replaces the need for separate H&E staining and immunofluorescence staining of different tissue sections, reducing overall processing time and complexity.
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 accurate pathology assessment by generating a simulated H&E image that does not interfere with subsequent multiplexed fluorescence imaging, preserving sample integrity and facilitating efficient triage and analysis.
Implementation Method 1
measuring information corresponding to one or more stains of the first stain composition in the biological sample, where the one or more stains comprise a fluorescent counterstain
Implementation Method 2
measuring information corresponding to one or more stains of the second stain composition in the biological sample, where the one or more stains comprise eosin and/or indigo carmine
Data Source
AI summary
Methods include applying a first stain composition comprising a fluorescent counterstain to a biological sample, measuring information corresponding to one or more stains of the first stain composition, removing the fluorescent counterstain from the biological sample, applying a second stain composition to the biological sample, measuring information corresponding to one or more stains of the second stain composition in the biological sample, where the one or more stains include a fluorescent label, and generating a first image of the biological sample, where the first image corresponds to a pattern of simulated hematoxylin staining in the biological sample.