Exponential Fluorescent Amplification of Individual Nucleic Acids Using Clampfish Probes | AIChE

Other Sites & Tools

Technical Groups

Exponential Fluorescent Amplification of Individual Nucleic Acids Using Clampfish Probes

Authors 

Rouhanifard, S. H. - Presenter, University of Pennsylvania
Mellis, I. A., University of Pennsylvania
Dunagin, M., University of Pennsylvania
Bayatpour, S., University of Pennsylvania
Jiang, C. L., University of Pennsylvania
Dardani, I., University of Pennsylvania
Symmons, O., University of Pennsylvania
Emert, B., University of Pennsylvania
Torre, E., University of Pennsylvania
Cote, A., University of Pennsylvania
Sullivan, A., Altius Institute
Stamatoyannopoulos, J., Altius Institute
Raj, A., University of Pennsylvania
Non-enzymatic, high-gain signal amplification methods with single-cell, single-molecule resolution are in great need. We present click-amplifying FISH (clampFISH) for the fluorescent detection of nucleic acids that combines the specificity of oligonucleotides with bioorthogonal click chemistry in order to achieve high specificity and extremely high-gain (>400x) signal amplification in single cells. We show that clampFISH signal enables detection of RNA species with low magnification microscopy and separation of cells by RNA levels via flow cytometry. Additionally, we show that the modular design of clampFISH probes enables multiplexing of RNA and DNA, that the locking mechanism prevents probe detachment in expansion microscopy, and that clampFISH can be applied to tissue samples.