Alternating laser excitation for solution-based single-molecule FRET

Achillefs Kapanidis, Devdoot Majumdar, Mike Heilemann, Eyal Nir, Shimon Weiss

Research output: Contribution to journalArticlepeer-review

10 Scopus citations


Single-molecule fluorescence resonance energy transfer (smFRET) has been widely applied to the study of fluorescently labeled biomolecules on surfaces and in solution. Sorting single molecules based on fluorescent dye stoichiometry provides one with further layers of information and also enables “filtering” of unwanted molecules from the analysis. We accomplish this sorting by using alternating laser excitation (ALEX) in combination with smFRET measurements; here we describe the implementation of these methodologies for the study of biomolecules in solution.

Original languageEnglish
Pages (from-to)979-987
Number of pages9
JournalCold Spring Harbor Protocols
Issue number11
StatePublished - 1 Nov 2015

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology


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