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dc.contributor.authorMitchell, C.A.
dc.contributor.authorKalies, S.
dc.contributor.authorCizmár, T.
dc.contributor.authorHeisterkamp, A.
dc.contributor.authorTorrance, L.
dc.contributor.authorRoberts, A.G.
dc.contributor.authorGunn-Moore, F.J.
dc.contributor.authorDholakia, K.
dc.date.accessioned2013-12-02T10:31:02Z
dc.date.available2013-12-02T10:31:02Z
dc.date.issued2013-11-14
dc.identifier.citationMitchell , C A , Kalies , S , Cizmár , T , Heisterkamp , A , Torrance , L , Roberts , A G , Gunn-Moore , F J & Dholakia , K 2013 , ' Femtosecond optoinjection of intact tobacco BY-2 cells using a reconfigurable photoporation platform ' PLoS One , vol 8 , no. 11 , e79235 . DOI: 10.1371/journal.pone.0079235en
dc.identifier.issn1932-6203
dc.identifier.otherPURE: 71205869
dc.identifier.otherPURE UUID: eae2748c-d5e9-4cb7-92b4-718acd4e1100
dc.identifier.otherScopus: 84893643628
dc.identifier.otherScopus: 84893643628
dc.identifier.urihttp://hdl.handle.net/10023/4234
dc.description.abstractA tightly-focused ultrashort pulsed laser beam incident upon a cell membrane has previously been shown to transiently increase cell membrane permeability while maintaining the viability of the cell, a technique known as photoporation. This permeability can be used to aid the passage of membrane-impermeable biologically-relevant substances such as dyes, proteins and nucleic acids into the cell. Ultrashort-pulsed lasers have proven to be indispensable for photoporating mammalian cells but they have rarely been applied to plant cells due to their larger sizes and rigid and thick cell walls, which significantly hinders the intracellular delivery of exogenous substances. Here we demonstrate and quantify femtosecond optical injection of membrane impermeable dyes into intact BY-2 tobacco plant cells growing in culture, investigating both optical and biological parameters. Specifically, we show that the long axial extent of a propagation invariant (“diffraction-free”) Bessel beam, which relaxes the requirements for tight focusing on the cell membrane, outperforms a standard Gaussian photoporation beam, achieving up to 70% optoinjection efficiency. Studies on the osmotic effects of culture media show that a hypertonic extracellular medium was found to be necessary to reduce turgor pressure and facilitate molecular entry into the cells.en
dc.format.extent10en
dc.language.isoeng
dc.relation.ispartofPLoS Oneen
dc.rights© 2013 Mitchell et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en
dc.subjectPulsed laser beamen
dc.subjectCell membrane permeabilityen
dc.subjectPhotoporationen
dc.subjectFemtosecond optical injectionen
dc.subjectTobacco BY-2 cellsen
dc.subjectQ Scienceen
dc.subject.lccQen
dc.titleFemtosecond optoinjection of intact tobacco BY-2 cells using a reconfigurable photoporation platformen
dc.typeJournal articleen
dc.description.versionPublisher PDFen
dc.contributor.institutionUniversity of St Andrews. School of Biologyen
dc.contributor.institutionUniversity of St Andrews. The University of St Andrewsen
dc.contributor.institutionUniversity of St Andrews. School of Physics and Astronomyen
dc.contributor.institutionUniversity of St Andrews. School of Medicineen
dc.contributor.institutionUniversity of St Andrews. Geography & Sustainable Developmenten
dc.contributor.institutionUniversity of St Andrews. Institute of Behavioural and Neural Sciencesen
dc.contributor.institutionUniversity of St Andrews. Biomedical Sciences Research Complexen
dc.identifier.doihttps://doi.org/10.1371/journal.pone.0079235
dc.description.statusPeer revieweden


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