Composite Imaging of Protein and Nucleic Acid Targets
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Solution Overview
Problem
Current methods for detecting protein expression and target nucleic acid sequences in biological samples often require multiple samples and cannot provide comprehensive, cell-by-cell analysis, limiting the ability to determine relative characteristics such as presence, absence, concentration, and spatial distribution of multiple biological targets.
Innovation Solution
A method for generating composite images of biological samples that combines fluorescent detection of protein expression and target nucleic acid sequences on the same section of the sample, allowing for the relative locations of both targets to be visualized and analyzed.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of information
If multiple separate analysis methods are used to detect different targets, then comprehensive target characterization is achieved, but multiple biological samples are required and spatial correlation is lost
Solution Approach 1:
The patent combines multiple detection methods (immunofluorescence for protein detection and FISH for nucleic acid detection) into a single integrated assay performed on one biological sample. This merging allows simultaneous detection of multiple targets while preserving spatial correlation information and reducing the number of samples required.
Solution Approach 2:
The invention creates a universal detection platform that can detect both protein and nucleic acid targets using a single sample. The method employs universal sample preparation procedures and a unified imaging approach that accommodates multiple target types, making the system multi-functional rather than requiring separate specialized assays.
2Measurement precision
If fluorescent detection system is used for protein expression analysis, then sensitivity is improved, but the number of detectable targets is limited
Solution Approach 1:
The patent utilizes spectral parameter differentiation by selecting fluorophores with distinct emission spectra that can be resolved by the detection system. By carefully choosing fluorophores with non-overlapping emission wavelengths and adjusting detection parameters, the system can detect multiple targets simultaneously while maintaining high sensitivity for each target.
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 comprehensive, cell-by-cell analysis of protein and nucleic acid expression, improving the accuracy of disease characterization and treatment decision-making by providing relative spatial information of multiple targets in a single biological sample.
Implementation Method 1
detecting, by fluorescence, signals from the first binder and the fluorescent marker
Implementation Method 2
contacting the same sample from step (1) with a probe for each of at least one target nucleic acid sequence thus hybridizing the probes with the target nucleic acid sequence
Data Source
AI summary
A method for providing a composite image of a single biological sample, comprising the steps of generating a first image of the biological sample with a target protein, generating a second image of the biological sample with a target nucleic acid sequence, and generating a composite image that provides the relative locations of both the target protein and the target nucleic acid sequence. Also provided is a method of analyzing a biological sample, comprising providing a composite image of the biological sample according to the method for providing a composite image, and analyzing the expression of the protein and the nucleic acid sequences of interest from the composite image. Further provided are systems and kits that comprise the means for executing the novel methods.