Temporally resolved direct delivery of second messengers into cells using nanostraws
Abstract
Second messengers are biomolecules with the critical role of conveying information to intracellular targets. They are typically membrane-impermeable and only enter cells through tightly regulated transporters. Current methods for manipulating second messengers in cells require preparation of modified cell lines or significant disruptions in cell function, especially at the cell membrane. Here we demonstrate that 100 nm diameter 'nanostraws' penetrate the cell membrane to directly modulate second messenger concentrations within cells. Nanostraws are hollow vertical nanowires that provide a fluidic conduit into cells to allow time-resolved delivery of the signaling ion Ca^(2+) without chemical permeabilization or genetic modification, minimizing cell perturbation. By integrating the nanostraw platform into a microfluidic device, we demonstrate coordinated delivery of Ca^(2+) ions into hundreds of cells at the time scale of several seconds with the ability to deliver complex signal patterns, such as oscillations over time. The diffusive nature of nanostraw delivery gives the platform unique versatility, opening the possibility for time-resolved delivery of any freely diffusing molecules.
Additional Information
© 2016 The Royal Society of Chemistry. Received 7th April 2016, Accepted 1st June 2016. First published online 02 Jun 2016. We would like to thank the David Goldhaber-Gordon and Yi Cui labs for use of equipment and the Sarah Heilshorn lab for use of microscopes. Additional experiments were performed at the Stanford Nanofabrication Facility and the Stanford Nano Shared Facilities. This project was supported by a Bio-X Interdisciplinary Initiatives Project (IIP), A. M. X. was supported by NSF and NDSEG fellowships. S. A. K. was supported by a fellowship from the Phelan-McDermid Syndrome Foundation.Attached Files
Published - c6lc00463f.pdf
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Additional details
- Eprint ID
- 68509
- Resolver ID
- CaltechAUTHORS:20160620-082156896
- Bio-X Interdisciplinary Initiatives Project (IIP)
- NSF
- National Defense Science and Engineering Graduate (NDSEG) Fellowship
- Phelan-McDermid Syndrome Foundation
- Created
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2016-06-20Created from EPrint's datestamp field
- Updated
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2021-11-11Created from EPrint's last_modified field