Femtosecond optical transfection of cells: viability and efficiency.
Photoporation is a rapidly expanding technique for the introduction of macromolecules into single cells. However, there remains no study into the true efficiency of this procedure. Here, we present a detailed analysis of transfection efficiency and cell viability for femtosecond optical transfection using a titanium sapphire laser at 800 nm. Photoporation of 4000 Chinese Hamster ovary cells was performed, representing the largest optical transfection study reported to date. We have investigated a range of laser fluences at the cell membrane and, at 1.2 microJ/cm(2), have found an average transfection efficiency of 50 +/- 10%. Contrary to recent literature, in which 100% efficiency is claimed, our measure of efficiency accounts for all irradiated cells, including those lost as a result of laser treatment, thereby providing a true biological measure of the technique.
Stevenson
D
D
Interdisciplinary Centre for Medical Photonics, School of Physics & Astronomy, University of St Andrews, St Andrews, KY16 9SS, Scotland, UK. ds50@st-andrews.ac.uk
Agate
B
B
Tsampoula
X
X
Fischer
P
P
Brown
C T A
CT
Sibbett
W
W
Riches
A
A
Gunn-Moore
F
F
Dholakia
K
K
D000818:Animals; D002470:Cell Survival / Q000502:physiology*; D006224:Cricetinae; D005260:Female; D054023:Lasers, Semiconductor; D055620:Optical Phenomena; D052898:Optical Tweezers; D010053:Ovary / Q000166:cytology*; D014162:Transfection / Q000379:methods*