Fluorescent probes and bioconjugation chemistries for single-molecule fluorescence analysis of biomolecules

Achillefs N. Kapanidis, Shimon Weiss

Research output: Contribution to journalArticlepeer-review

131 Scopus citations

Abstract

A popular method for probing single molecules is single-molecule fluorescence spectroscopy (SMFS). SMFS has allowed striking visualizations of single biomolecules at work, including the rotation of the rotor subunit of the rotary stepper motor protein F1-ATPase, the sliding of kinesin on microtubules, the translocation of RNA polymerase (RNAP) molecules on double-stranded DNA, the formation of ribozyme intermediates, and the real-time infection of a cell by single virus particles. This paper describes the properties of SMFS-compatible fluorophores, list considerations for popular assays, provides examples of site-specific labeling strategies that enabled SMFS observations, and looks at fluorophores and labeling strategies that can answer future challenges.

Original languageEnglish
Pages (from-to)10953-10964
Number of pages12
JournalJournal of Chemical Physics
Volume117
Issue number24
DOIs
StatePublished - 22 Dec 2002
Externally publishedYes

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