Chromogen Layering for Multiplex Analyte Detection
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Solution Overview
Problem
Current methods for staining biological samples can only determine the presence or absence of a single analyte, limiting the ability to identify multiple analytes simultaneously within a single sample.
Innovation Solution
The chromogen layering method involves using multiple chromogens and antibodies or probes to generate distinct colors for different analytes, allowing for the identification of multiple analytes by overlapping colors, enabling the simultaneous detection of two or more analytes on a single specimen.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If a single chromogen and stain are used for staining, then the staining process is simple and straightforward, but only a single analyte can be determined
Solution Approach 1:
The staining process is segmented into multiple sequential steps, each applying a different chromogen layer. The first chromogen layer stains for a first analyte, the second chromogen layer stains for a second analyte, and so on. This segmentation allows multiple analytes to be identified simultaneously while maintaining a systematic and manageable process flow.
Solution Approach 2:
The invention adds a temporal dimension to the staining process by applying chromogens sequentially rather than simultaneously. Each chromogen layer is applied in a specific sequence, with intermediate processing steps between layers. This dimensional approach to process organization enables multiplex detection while maintaining process control.
2Loss of information
If multiple chromogens and stains are applied sequentially, then multiple analytes can be identified simultaneously, but the staining process becomes more complex
Solution Approach 1:
The chromogens are selected and prepared in advance with specific color properties in mind. The first chromogen is chosen to produce a base color, and subsequent chromogens are selected to produce colors that will create distinguishable combinations when layered. This preliminary selection and preparation reduces complexity during the actual staining process.
Solution Approach 2:
The invention utilizes color changes and color mixing principles to encode multiple analyte detections. Different chromogen combinations produce distinct color outcomes (e.g., yellow + blue = green, red + blue = purple), allowing multiple analytes to be distinguished by their color signatures. This color-based encoding system efficiently transmits multiple pieces of information simultaneously.
3Measurement precision
If chromogens are overlaid to produce unique colors, then diagnostic accuracy improves, but the process requires precise control
Solution Approach 1:
The invention controls the staining process by adjusting key parameters including chromogen concentration, application time, and layering sequence. Each chromogen layer is applied with specific parameters optimized for that layer, allowing precise control over the final color outcome while maintaining ease of operation through standardized parameter sets.
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 method allows for the identification of multiple analytes in a single sample by producing unique colors, expanding the capability to detect and differentiate between various analytes, thereby improving diagnostic accuracy.
Implementation Method 1
contacting the section with a first chromogen that reacts with the first marker to generate a first color
Data Source
AI summary
The present disclosure relates methods of chromogen layering, wherein a first chromogen and a first stain (color) are produced on a sample, specific for a first analyte, followed by a second chromogen and a second stain (color) being produced on the same sample, specific for a second analyte. In addition, if desired, by overlaying the second stain on top of the first stain, a unique third color is produced that is specific for a third analyte. Therefore, the distribution of different colors throughout the sample could be used to identify at least two or more analytes simultaneously within a single sample.