Femtosecond Optoinjection of Intact Tobacco BY-2 Cells Using a Reconfigurable Photoporation Platform

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dc.identifier.uri http://dx.doi.org/10.15488/3505
dc.identifier.uri http://www.repo.uni-hannover.de:8080/handle/123456789/3535
dc.contributor.author Mitchell, Claire A.
dc.contributor.author Kalies, Stefan
dc.contributor.author Cizmar, Tomas
dc.contributor.author Heisterkamp, Alexander
dc.contributor.author Torrance, Lesley
dc.contributor.author Roberts, Alison G.
dc.contributor.author Gunn-Moore, Frank J.
dc.contributor.author Dholakia, Kishan
dc.date.accessioned 2018-07-03T11:45:28Z
dc.date.available 2018-07-03T11:45:28Z
dc.date.issued 2013
dc.identifier.citation Mitchell, Claire A.; Kalies, Stefan; Cizmar, Tomas; Heisterkamp, Alexander; Torrance, Lesley et al.: Femtosecond Optoinjection of Intact Tobacco BY-2 Cells Using a Reconfigurable Photoporation Platform. In: PLoS One 8 (2013), Nr. 11, e79235. DOI: https://doi.org/10.1371/journal.pone.0079235
dc.description.abstract A 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. eng
dc.language.iso eng
dc.publisher San Francisco, CA : Public Library of Science
dc.relation.ispartofseries PLoS One 8 (2013), Nr. 11
dc.rights CC BY 4.0 Unported
dc.rights.uri https://creativecommons.org/licenses/by/4.0/
dc.subject plant-cells eng
dc.subject optical transfection eng
dc.subject mammalian-cells eng
dc.subject laser-pulses eng
dc.subject gene-transfer eng
dc.subject delivery eng
dc.subject single eng
dc.subject proteins eng
dc.subject microinjection eng
dc.subject expression eng
dc.subject.ddc 500 | Naturwissenschaften ger
dc.subject.ddc 610 | Medizin, Gesundheit ger
dc.title Femtosecond Optoinjection of Intact Tobacco BY-2 Cells Using a Reconfigurable Photoporation Platform eng
dc.type Article
dc.type Text
dc.relation.essn 1932-6203
dc.relation.doi https://doi.org/10.1371/journal.pone.0079235
dc.bibliographicCitation.issue 11
dc.bibliographicCitation.volume 8
dc.bibliographicCitation.firstPage e79235
dc.description.version publishedVersion
tib.accessRights frei zug�nglich

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