dc.contributor.author |
Baddeley, David |
en |
dc.contributor.author |
Crossman, David |
en |
dc.contributor.author |
Rossberger, S |
en |
dc.contributor.author |
Cheyne, Juliette |
en |
dc.contributor.author |
Montgomery, Johanna |
en |
dc.contributor.author |
Jayasinghe, ID |
en |
dc.contributor.author |
Cremer, C |
en |
dc.contributor.author |
Cannell, MB |
en |
dc.contributor.author |
Soeller, C |
en |
dc.date.accessioned |
2012-03-25T20:41:05Z |
en |
dc.date.issued |
2011 |
en |
dc.identifier.citation |
PLoS One 6(5):1-10 Article number e20645 2011 |
en |
dc.identifier.issn |
1932-6203 |
en |
dc.identifier.uri |
http://hdl.handle.net/2292/15212 |
en |
dc.description.abstract |
Background: Optical super-resolution imaging of fluorescently stained biological samples is rapidly becoming an important tool to investigate protein distribution at the molecular scale. It is therefore important to develop practical super-resolution methods that allow capturing the full three-dimensional nature of biological systems and also can visualize multiple protein species in the same sample. Methodology/Principal Findings: We show that the use of a combination of conventional near-infrared dyes, such as Alexa 647, Alexa 680 and Alexa 750, all excited with a 671 nm diode laser, enables 3D multi-colour super-resolution imaging of complex biological samples. Optically thick samples, including human tissue sections, cardiac rat myocytes and densely grown neuronal cultures were imaged with lateral resolutions of ,15 nm (std. dev.) while reducing marker cross-talk to ,1%. Using astigmatism an axial resolution of ,65 nm (std. dev.) was routinely achieved. The number of marker species that can be distinguished depends on the mean photon number of single molecule events. With the typical photon yields from Alexa 680 of ,2000 up to 5 markers may in principle be resolved with ,2% crosstalk. Conclusions/Significance: Our approach is based entirely on the use of conventional, commercially available markers and requires only a single laser. It provides a very straightforward way to investigate biological samples at the nanometre scale and should help establish practical 4D super-resolution microscopy as a routine research tool in many laboratories. |
en |
dc.publisher |
The Authors |
en |
dc.relation.ispartofseries |
PLOS ONE |
en |
dc.rights |
Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher.
Details obtained from http://www.sherpa.ac.uk/romeo/issn/1932-6203/ |
en |
dc.rights.uri |
https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm |
en |
dc.title |
4D super-resolution microscopy with conventional fluorophores and single wavelength excitation in optically thick cells and tissues |
en |
dc.type |
Journal Article |
en |
dc.identifier.doi |
10.1371/journal.pone.0020645 |
en |
pubs.issue |
5 |
en |
pubs.begin-page |
1 |
en |
pubs.volume |
6 |
en |
dc.rights.holder |
Copyright: The Authors |
en |
dc.identifier.pmid |
21655189 |
en |
pubs.end-page |
10 |
en |
pubs.publication-status |
Published |
en |
dc.rights.accessrights |
http://purl.org/eprint/accessRights/RestrictedAccess |
en |
pubs.subtype |
Article |
en |
pubs.elements-id |
257334 |
en |
pubs.org-id |
Bioengineering Institute |
en |
pubs.org-id |
Medical and Health Sciences |
en |
pubs.org-id |
Medical Sciences |
en |
pubs.org-id |
Physiology Division |
en |
pubs.org-id |
Science |
en |
pubs.org-id |
Science Research |
en |
pubs.org-id |
Maurice Wilkins Centre (2010-2014) |
en |
pubs.number |
e20645 |
en |
pubs.record-created-at-source-date |
2011-12-08 |
en |
pubs.dimensions-id |
21655189 |
en |