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Background:
Single-molecule fluorescent in situ hybridization (smFISH) stands as the gold standard for RNA visualization and quantification in cellular studies. However, its application in high-throughput methods like flow cytometry or single-cell RNA sequencing faces significant challenges due to limited signal intensity and nuclear accessibility constraints. The nuclear envelope and chromatin structure create barriers that hinder effective probe penetration and binding, reducing the sensitivity of RNA detection methods. Current approaches often require extensive optimization and may not provide sufficient signal-to-noise ratios for reliable downstream applications, particularly when analyzing rare transcripts or conducting large-scale cellular screening.
Technical Overview:
Northeastern researchers have developed a novel method that overcomes the limitations of current nuclear RNA amplification techniques. The approach centers on optimizing nuclear accessibility through controlled permeabilization protocols combined with enhanced signal amplification strategies. The method employs specific probe designs and amplification cascades that dramatically increase signal intensity while preserving cellular morphology and RNA integrity. This technique enables robust detection of nuclear RNAs with significantly improved sensitivity compared to standard smFISH protocols. The system has been optimized for compatibility with flow cytometry platforms, enabling high-throughput single-cell analysis based on nuclear RNA expression profiles.
Benefits:
- Enhanced accessibility of probes to nuclear RNAs
- Signal enhancement for nuclear RNAs previously difficult to detect
- Compatibility with high-throughput screening platforms
- Preserved cellular integrity during analysis
- Improved signal-to-noise ratios for rare transcript detection
Application:
- Single-Cell Imaging: Advanced profiling of nuclear RNA expression at individual cell level
- Flow Cytometry: High-throughput cell sorting based on nuclear RNA expression
- Drug Screening: Identifying compounds that affect nuclear RNA processing
- Disease Research: Studying nuclear RNA dysfunction in various pathologies
Opportunity:
- License
- Research collaboration
