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Proceedings Paper

Femtosecond cellular transfection using a non-diffracting beam
Author(s): X. Tsampoula; V. Garcés-Chávez; M. Comrie; D. J. Stevenson; B. Agate; C. T. A. Brown; F. Gunn-Moore; K. Dholakia
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Paper 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.

Paper Details

Date Published: 20 February 2008
PDF: 10 pages
Proc. SPIE 6854, Optical Interactions with Tissue and Cells XIX, 68541L (20 February 2008); doi: 10.1117/12.762677
Show Author Affiliations
X. Tsampoula, Univ. of St. Andrews (United Kingdom)
V. Garcés-Chávez, Univ. of St. Andrews (United Kingdom)
M. Comrie, Univ. of St. Andrews (United Kingdom)
D. J. Stevenson, Univ. of St. Andrews (United Kingdom)
B. Agate, Univ. of St. Andrews (United Kingdom)
C. T. A. Brown, Univ. of St. Andrews (United Kingdom)
F. Gunn-Moore, Univ. of St. Andrews (United Kingdom)
K. Dholakia, Univ. of St. Andrews (United Kingdom)

Published in SPIE Proceedings Vol. 6854:
Optical Interactions with Tissue and Cells XIX
Steven L. Jacques; William P. Roach; Robert J. Thomas, Editor(s)

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