Journal
EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS
Volume 36, Issue 6, Pages 669-674Publisher
SPRINGER
DOI: 10.1007/s00249-007-0133-7
Keywords
single-molecule fluorescence spectroscopy; alternating-laser excitation (ALEX); Forster resonance energy transfer (FRET); biomolecular interactions
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Funding
- EPSRC [EP/D058775/1] Funding Source: UKRI
- Engineering and Physical Sciences Research Council [EP/D058775/1] Funding Source: researchfish
- NIGMS NIH HHS [R01 GM069709, GM069709-01, R01 GM069709-01A1] Funding Source: Medline
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Alternating-laser excitation (ALEX) spectroscopy has recently been added to the single-molecule spectroscopy toolkit. ALEX monitors interaction and stoichiometry of biomolecules, reports on biomolecular structure by measuring accurate Forster resonance energy transfer (FRET) efficiencies, and allows sorting of subpopulations on the basis of stoichiometry and FRET. Here, we demonstrate that a simple combination of one continuous-wave donor-excitation laser and one directly modulated acceptor-excitation laser (Periodic Acceptor eXcitation) is sufficient to recapitulate the capabilities of ALEX while minimizing the cost and complexity associated with use of modulation techniques.
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