Chau, C. orcid.org/0000-0002-3134-6798, Maffeo, C. orcid.org/0000-0001-9927-1502, Aksimentiev, A. orcid.org/0000-0002-6042-8442 et al. (3 more authors) (2024) Single molecule delivery into living cells. Nature Communications, 15. 4403. ISSN 2041-1723
Abstract
Controlled manipulation of cultured cells by delivery of exogenous macromolecules is a cornerstone of experimental biology. Here we describe a platform that uses nanopipettes to deliver defined numbers of macromolecules into cultured cell lines and primary cells at single molecule resolution. In the nanoinjection platform, the nanopipette is used as both a scanning ion conductance microscope (SICM) probe and an injection probe. The SICM is used to position the nanopipette above the cell surface before the nanopipette is inserted into the cell into a defined location and to a predefined depth. We demonstrate that the nanoinjection platform enables the quantitative delivery of DNA, globular proteins, and protein fibrils into cells with single molecule resolution and that delivery results in a phenotypic change in the cell that depends on the identity of the molecules introduced. Using experiments and computational modeling, we also show that macromolecular crowding in the cell increases the signal-to-noise ratio for the detection of translocation events, thus the cell itself enhances the detection of the molecules delivered.
Metadata
Item Type: | Article |
---|---|
Authors/Creators: |
|
Copyright, Publisher and Additional Information: | © The Author(s) 2024. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/ licenses/by/4.0/. |
Dates: |
|
Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Electronic & Electrical Engineering (Leeds) > Pollard Institute (Leeds) |
Funding Information: | Funder Grant number EPSRC (Engineering and Physical Sciences Research Council) EP/S01764X/1 EPSRC (Engineering and Physical Sciences Research Council) EP/W004933/1 BBSRC (Biotechnology & Biological Sciences Research Council) Not Known Royal Society RG160273 |
Depositing User: | Symplectic Publications |
Date Deposited: | 16 May 2024 09:05 |
Last Modified: | 23 Jan 2025 15:43 |
Published Version: | https://www.nature.com/articles/s41467-024-48608-3 |
Status: | Published |
Publisher: | Nature Portfolio |
Identification Number: | 10.1038/s41467-024-48608-3 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:212552 |
Download
Filename: Single molecule delivery into living cells.pdf
Licence: CC-BY 4.0