Strategies for Overcoming the Single-Molecule Concentration Barrier

David S. White, Mackinsey A. Smith, Baron Chanda, Randall H. Goldsmith

Research output: Contribution to journalReview articlepeer-review

3 Scopus citations

Abstract

Fluorescence-based single-molecule approaches have helped revolutionize our understanding of chemical and biological mechanisms. Unfortunately, these methods are only suitable at low concentrations of fluorescent molecules so that single fluorescent species of interest can be successfully resolved beyond background signal. The application of these techniques has therefore been limited to high-affinity interactions despite most biological and chemical processes occurring at much higher reactant concentrations. Fortunately, recent methodological advances have demonstrated that this concentration barrier can indeed be broken, with techniques reaching concentrations as high as 1 mM. The goal of this Review is to discuss the challenges in performing single-molecule fluorescence techniques at high-concentration, offer applications in both biology and chemistry, and highlight the major milestones that shatter the concentration barrier. We also hope to inspire the widespread use of these techniques so we can begin exploring the new physical phenomena lying beyond this barrier.

Original languageEnglish
Pages (from-to)239-257
Number of pages19
JournalACS Measurement Science Au
Volume3
Issue number4
DOIs
StatePublished - Aug 16 2023

Keywords

  • FRET
  • Single-molecule
  • dynamics
  • fluorescence
  • fluorogenic
  • high-concentration
  • spectroscopy
  • zero mode waveguide

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