Femtosecond cellular transfection using a non-diffracting beam

X. Tsampoula, V. Garcés-Chávez, M. Comrie, D. J. Stevenson, B. Agate, C. T.A. Brown, F. Gunn-Moore, K. Dholakia

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Efficient DNA delivery into single living cells would be a very powerful capability for cell biologists for elucidating basic cellular functions but also in other fields such as applied drug discovery and gene therapy. The ability to gently permeate the cell membrane and introduce foreign DNA with the assistance of lasers is a powerful methodology but requires exact focusing due to the required two-photon power density. Here, we demonstrate a laser-mediated delivery method of the red fluorescent protein DS-RED into Chinese hamster Ovary (CHO) cells. We used an elongated beam of light created by a Bessel beam (BB) which obviates the need to locate precisely the cell membrane, permitting two-photon excitation along a line leading to cell transfection. Assuming a threshold for transfection of 20%, the BB gives us transfection over twenty times the axial distance compared to the Gaussian beam of equivalent core diameter. In addition, by exploiting the BB property of reconstruction, we demonstrate successful transfection of CHO cells which involves the BB passing through an obstructive layer and re forming itself prior to reaching the cell membrane. In the light of this exciting result, one can envisage the possibility of achieving transfection through multiple cell monolayer planes and tissues using this novel light field, eliminating this way the stringent requirements for tight focusing.

Original languageEnglish
Title of host publicationOptical Interactions with Tissue and Cells XIX
DOIs
Publication statusPublished - 2008
EventOptical Interactions with Tissue and Cells XIX - San Jose, CA, United States
Duration: 2008 Jan 212008 Jan 23

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume6854
ISSN (Print)1605-7422

Conference

ConferenceOptical Interactions with Tissue and Cells XIX
CountryUnited States
CitySan Jose, CA
Period08/1/2108/1/23

All Science Journal Classification (ASJC) codes

  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Atomic and Molecular Physics, and Optics
  • Radiology Nuclear Medicine and imaging

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  • Cite this

    Tsampoula, X., Garcés-Chávez, V., Comrie, M., Stevenson, D. J., Agate, B., Brown, C. T. A., Gunn-Moore, F., & Dholakia, K. (2008). Femtosecond cellular transfection using a non-diffracting beam. In Optical Interactions with Tissue and Cells XIX [68541L] (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 6854). https://doi.org/10.1117/12.762677