LANTERN Probe Cascade for Multiplexed Transcriptomic Imaging
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Solution Overview
Problem
Current methods for transcriptomic profiling, such as qPCR and next-generation sequencing, lack single molecule sensitivity and are time-consuming and costly, while imaging techniques are limited to quantifying a few hundred genes, failing to provide comprehensive gene expression analysis efficiently.
Innovation Solution
The Linked Amplification Tethered with Exponential Radiance (LANTERN) method uses a cascade of primary, secondary, tertiary, and quaternary probes for exponential signal amplification in multiplexed imaging, allowing for precise and deterministic amplification of amplicons, enabling the profiling of thousands of genes with high accuracy and efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If imaging techniques are used for mRNA transcripts quantification, then single molecule sensitivity is achieved, but the number of genes that can be quantified is limited to a few hundred
Solution Approach 1:
The invention segments the gene quantification process into multiple sequential imaging rounds, where different subsets of genes are detected in each round. By dividing the transcriptome into manageable segments that can be imaged separately and then integrated computationally, the system overcomes the physical limitation of detecting only a few hundred genes simultaneously while maintaining single-molecule sensitivity throughout.
Solution Approach 2:
The invention employs periodic action through sequential imaging rounds, where the same sample is imaged multiple times with different probe sets. Each round detects a specific subset of genes, and the process is repeated with new probes to capture additional genes. This periodic re-imaging of the same sample enables comprehensive transcriptome-wide quantification while preserving single-molecule detection capability.
2Adaptability or versatility
If qPCR and next generation sequencing are used for transcriptomic profiling, then comprehensive gene expression analysis is achieved, but single molecule sensitivity and time efficiency are lost
Solution Approach 1:
The invention uses copying by implementing multiple sequential imaging rounds that capture the same biological sample repeatedly. Each round produces a copy of the detection data for different gene subsets, which are then integrated to form a comprehensive transcriptome profile. This copying approach allows the system to achieve both comprehensive gene coverage and single-molecule sensitivity without requiring physical amplification that would compromise accuracy.
3Measurement precision
If sequential hybridization rounds are performed to amplify signal, then detection sensitivity is improved, but imaging time increases
Solution Approach 1:
The invention applies preliminary action by performing extensive signal amplification during the probe hybridization step itself. The probes are designed to bind with high affinity and generate strong signals that reduce the need for subsequent amplification rounds. By preparing the detection system in advance with optimized probes that provide built-in signal enhancement, the method minimizes the number of sequential imaging rounds required, thereby reducing total imaging time while maintaining high detection sensitivity.
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
LANTERN achieves scalable and accurate signal amplification, significantly reducing imaging time and cost, enabling the detection of thousands of genes with improved sensitivity and specificity compared to existing methods, while maintaining stability across multiple hybridization and imaging rounds.
Implementation Method 1
one or more primary probes capable of binding one or more targets
Implementation Method 2
each primary probe comprises one or more secondary probe binding sites
Implementation Method 3
one or more readout probes capable of binding to a binding site on the one or more primary, secondary, tertiary, or quaternary probes and capable of being detected
Data Source
AI summary
Disclosed herein is a composition for linked amplification tethered with exponential radiance for signal amplification. Also disclosed herein, is a kit for linked amplification tethered with exponential radiance for signal amplification. Also disclosed herein, is a method linked amplification tethered with exponential radiance for signal amplification.