Abstract
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 μJ/cm2, 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.
| Original language | English |
|---|---|
| Pages (from-to) | 7125-7133 |
| Number of pages | 9 |
| Journal | Optics Express |
| Volume | 14 |
| Issue number | 16 |
| DOIs | |
| Publication status | Published - 2006 Jan 1 |
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All Science Journal Classification (ASJC) codes
- Atomic and Molecular Physics, and Optics
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Femtosecond optical transfection of cells:viability and efficiency. / Stevenson, D.; Agate, B.; Tsampoula, X.; Fischer, P.; Brown, C. T.A.; Sibbett, W.; Riches, A.; Gunn-Moore, F.; Dholakia, K.
In: Optics Express, Vol. 14, No. 16, 01.01.2006, p. 7125-7133.Research output: Contribution to journal › Article
TY - JOUR
T1 - Femtosecond optical transfection of cells:viability and efficiency
AU - Stevenson, D.
AU - Agate, B.
AU - Tsampoula, X.
AU - Fischer, P.
AU - Brown, C. T.A.
AU - Sibbett, W.
AU - Riches, A.
AU - Gunn-Moore, F.
AU - Dholakia, K.
PY - 2006/1/1
Y1 - 2006/1/1
N2 - 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 μJ/cm2, 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.
AB - 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 μJ/cm2, 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.
UR - http://www.scopus.com/inward/record.url?scp=33746924833&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33746924833&partnerID=8YFLogxK
U2 - 10.1364/OE.14.007125
DO - 10.1364/OE.14.007125
M3 - Article
C2 - 19529083
AN - SCOPUS:33746924833
VL - 14
SP - 7125
EP - 7133
JO - Optics Express
JF - Optics Express
SN - 1094-4087
IS - 16
ER -