Characterization and application of controllable local chemical changes produced by reagent delivery from a nanopipet

Joe D. Piper, Chao Li, Chien Jung Lo, Richard Berry, Yuri Korchev, Liming Ying, David Klenerman

Research output: Contribution to journalArticlepeer-review

35 Scopus citations

Abstract

We introduce a versatile method that allows local and repeatable delivery (or depletion) of any water-soluble reagent from a nanopipet in ionic solution to make localized controlled changes in reagent concentration at a surface. In this work, Na+ or OH- ions were dosed from the pipet using pulsed voltage-driven delivery. Total internal reflection fluorescence from CoroNa Green dye in the bath for Na+ ions or fluorescein in the bath for pH quantified the resulting changes in local surface concentration. These changes had a time response as short as 10 ms and a radius of 1-30 μm and depended on the diameter of the pipet used, the applied voltage, and the pipet-surface separation. After the pipet dosing was characterized in detail, two proof-of-concept experiments on single cells and single molecules were then performed. We demonstrated local control of the sodium-sensitive flagellar motor in single Escherichia coli chimera on the time scale of 1 s by dosing sodium and monitoring the rotation of a 1 μm diameter bead fixed to the flagellum. We also demonstrated triggered single-molecule unfolding by dosing acid from the pipet to locally melt individual molecules of duplex DNA, as observed using fluorescent resonance energy transfer.

Original languageEnglish
Pages (from-to)10386-10393
Number of pages8
JournalJournal of the American Chemical Society
Volume130
Issue number31
DOIs
StatePublished - 6 Aug 2008

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