Nanochannel-based single molecule recycling

John F. Lesoine, Prahnesh A. Venkataraman, Peter C. Maloney, Mark E. Dumont, Lukas Novotny

Research output: Contribution to journalArticlepeer-review

Abstract

We present a method for measuring the fluorescence from a single molecule hundreds of times without surface immobilization. The approach is based on the use of electroosmosis to repeatedly drive a single target molecule in a fused silica nanochannel through a stationary laser focus. Single molecule fluorescence detected during the transit time through the laser focus is used to repeatedly reverse the electrical potential controlling the flow direction. Our method does not rely on continuous observation and therefore is less susceptible to fluorescence blinking than existing fluorescence-based trapping schemes. The variation in the turnaround times can be used to measure the diffusion coefficient on a single molecule level. We demonstrate the ability to recycle both proteins and DNA in nanochannels and show that the procedure can be combined with single-pair Förster energy transfer. Nanochannel-based single molecule recycling holds promise for studying conformational dynamics on the same single molecule in solution and without surface tethering.

Original languageEnglish (US)
Pages (from-to)3273-3278
Number of pages6
JournalNano Letters
Volume12
Issue number6
DOIs
StatePublished - Jun 13 2012

Keywords

  • FRET
  • Single molecule detection
  • electroosmosis
  • nanofluidics
  • trapping

ASJC Scopus subject areas

  • Bioengineering
  • Chemistry(all)
  • Materials Science(all)
  • Condensed Matter Physics
  • Mechanical Engineering

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