Novel technique for sorting cells based on nuclear RNA expression that enhances detection sensitivity while maintaining high sensitivity
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.
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