Penetration of Cell Membranes and Synthetic Lipid Bilayers by Nanoprobes
| Autor: | Aman Thomas, Nicholas A. Melosh, Andreas T. Schaefer, Matthew R. Angle, Andrew Wang |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
Membranes
Materials science Surface Properties Cell Membrane Lipid Bilayers Biophysics Correction Nanotechnology Penetration (firestop) Microscopy Atomic Force Actins Nanostructures Cell membrane HEK293 Cells Membrane medicine.anatomical_structure Membrane protein Microscopy Electron Scanning medicine Membrane fluidity Humans Cytoskeleton Lipid bilayer Intracellular |
| Zdroj: | Biophysical Journal. 107:2091-2100 |
| ISSN: | 0006-3495 |
| Popis: | Nanoscale devices have been proposed as tools for measuring and controlling intracellular activity by providing electrical and/or chemical access to the cytosol. Unfortunately, nanostructures with diameters of 50–500 nm do not readily penetrate the cell membrane, and rationally optimizing nanoprobes for cell penetration requires real-time characterization methods that are capable of following the process of membrane penetration with nanometer resolution. Although extensive work has examined the rupture of supported synthetic lipid bilayers, little is known about the applicability of these model systems to living cell membranes with complex lipid compositions, cytoskeletal attachment, and membrane proteins. Here, we describe atomic force microscopy (AFM) membrane penetration experiments in two parallel systems: live HEK293 cells and stacks of synthetic lipid bilayers. By using the same probes in both systems, we were able to clearly identify membrane penetration in synthetic bilayers and compare these events with putative membrane penetration events in cells. We examined membrane penetration forces for three tip geometries and 18 chemical modifications of the probe surface, and in all cases the median forces required to penetrate cellular and synthetic lipid bilayers with nanoprobes were greater than 1 nN. The penetration force was sensitive to the probe's sharpness, but not its surface chemistry, and the force did not depend on cell surface or cytoskeletal properties, with cells and lipid stacks yielding similar forces. This systematic assessment of penetration under various mechanical and chemical conditions provides insights into nanoprobe-cell interactions and informs the design of future intracellular nanoprobes. |
| Databáze: | OpenAIRE |
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