title: Local delivery of molecules from a nanopipette for quantitative receptor mapping on live cells.
abstract: Using nanopipettes to locally deliver molecules to the surface of living cells could potentially open up studies of biological processes down to the level of single molecules. However, in order to achieve precise and quantitative local delivery it is essential to be able to determine the amount and distribution of the molecules being delivered. In this work, we investigate how the size of the nanopipette, the magnitude of the applied pressure or voltage, which drives the delivery, and the distance to the underlying surface influences the number and spatial distribution of the delivered molecules. Analytical expressions describing the delivery are derived and compared with the results from finite element simulations and experiments on delivery from a 100 nm nanopipette in bulk solution and to the surface of sensory neurons. We then developed a setup for rapid and quantitative delivery to multiple subcellular areas, delivering the molecule capsaicin to stimulate opening of Transient Receptor Potential Vanilloid subfamily member 1 (TRPV1) channels, membrane receptors involved in pain sensation. Overall, precise and quantitative delivery of molecules from nanopipettes has been demonstrated, opening up many applications in biology such as locally stimulating and mapping receptors on the surface of live cells.
authors.affiliation: Division of Medicine, Imperial College London , London W12 0NN, U.K.
authors.forename: Babak
authors.initials: B
authors.lastname: Babakinejad
authors.forename: Peter
authors.initials: P
authors.lastname: Jönsson
authors.forename: Ainara
authors.initials: A
authors.lastname: López Córdoba
authors.forename: Paolo
authors.initials: P
authors.lastname: Actis
authors.forename: Pavel
authors.initials: P
authors.lastname: Novak
authors.forename: Yasufumi
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Andrew
authors.initials: A
authors.lastname: Shevchuk
authors.forename: Uma
authors.initials: U
authors.lastname: Anand
authors.forename: Praveen
authors.initials: P
authors.lastname: Anand
authors.forename: Anna
authors.initials: A
authors.lastname: Drews
authors.forename: Antonio
authors.initials: A
authors.lastname: Ferrer-Montiel
authors.forename: David
authors.initials: D
authors.lastname: Klenerman
authors.forename: Yuri E
authors.initials: YE
authors.lastname: Korchev
mesh_terms: D000818:Animals; D002211:Capsaicin / Q000737:chemistry / Q000378:metabolism*; D002478:Cells, Cultured; D005727:Ganglia, Spinal / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D036103:Nanotechnology / Q000295:instrumentation*; D010316:Particle Size; D051381:Rats; D017207:Rats, Sprague-Dawley; D013499:Surface Properties; D050916:TRPV Cation Channels / Q000737:chemistry / Q000378:metabolism*
chemical_list: D050916:TRPV Cation Channels; C493922:Trpv1 protein, rat; D002211:Capsaicin