Local Delivery of Molecules from a Nanopipette for Quantitative Receptor Mapping on Live Cells

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• 作者：Babak Babakinejad ; Peter J枚nsson ; Ainara L贸pez C贸rdoba ; Paolo Actis ; Pavel Novak ; Yasufumi Takahashi ; Andrew Shevchuk ; Uma Anand ; Praveen Anand ; Anna Drews ; Antonio Ferrer-Montiel ; David Klenerman ; Yuri E. Korchev
• 刊名：Analytical Chemistry
• 出版年：2013
• 出版时间：October 1, 2013
• 年：2013
• 卷：85
• 期：19
• 页码：9333-9342
• 全文大小：494K
• 年卷期：v.85,no.19(October 1, 2013)
• ISSN：1520-6882

文摘

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.