Transfection of cells using a violet diode laser for photoporation

Ben Agate; Lynn Paterson; Tanya K. Lake; J. E. Morris; A. E. Carruthers; Christian T. A. Brown; Wilson Sibbett; Kishan Dholakia; Muriel Comrie; R. Ferguson; P. E. Bryant; Andrew C. Riches; Frank J. Gunn-Moore

doi:10.1109/CLEOE.2005.1568410

Transfection of cells using a violet diode laser for photoporation

Ben Agate, Lynn Paterson, Tanya K. Lake, J. E. Morris, A. E. Carruthers, Christian T. A. Brown, Wilson Sibbett, Kishan Dholakia, Muriel Comrie, R. Ferguson, P. E. Bryant, Andrew C. Riches, Frank J. Gunn-Moore

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

Abstract

The introduction and subsequent expression of foreign DNA inside living mammalian cells (transfection) is achieved by photoporation with a violet diode laser. We direct a compact 405 nm laser diode source into an inverted optical microscope configuration and expose cells to 0.3 mW for 40 ms. The localized optical power density of ~1200 MW/m2 is six orders of magnitude lower than that used in femtosecond photoporation (~104 TW/m2). The tightly focused laser beam (with a localised focal volume of ~10-19 m3 ) perforates the cell plasma membrane to allow uptake of plasmid DNA containing an antibiotic resistant gene as well as the green fluorescent protein (GFP) gene. Successfully transfected cells then expand into clonal groups which are used to create stable cell lines. The use of the violet diode laser offers a new and simple poration technique compatible with standard microscopes and is the simplest method of laser-assisted cell poration reported to date

Original language	English
Title of host publication	2005 Conference on Lasers and Electro-Optics Europe
Publisher	OSA Publishing
Pages	634
Number of pages	1
ISBN (Print)	0780389743, 9780780389748
DOIs	https://doi.org/10.1109/CLEOE.2005.1568410
Publication status	Published - 2005
Event	2005 Conference on Lasers and Elctro-Optics Europe - Munich, Germany Duration: 12 Jun 2005 → 17 Jun 2005

Conference

Conference	2005 Conference on Lasers and Elctro-Optics Europe
Country/Territory	Germany
City	Munich
Period	12/06/05 → 17/06/05

ASJC Scopus subject areas

Control and Systems Engineering
Electrical and Electronic Engineering

Access to Document

10.1109/CLEOE.2005.1568410

Cite this

Agate, B., Paterson, L., Lake, T. K., Morris, J. E., Carruthers, A. E., Brown, C. T. A., Sibbett, W., Dholakia, K., Comrie, M., Ferguson, R., Bryant, P. E., Riches, A. C., & Gunn-Moore, F. J. (2005). Transfection of cells using a violet diode laser for photoporation. In 2005 Conference on Lasers and Electro-Optics Europe (pp. 634). OSA Publishing. https://doi.org/10.1109/CLEOE.2005.1568410

@inproceedings{b58e5974c93748a396e22d81ce42aa52,

title = "Transfection of cells using a violet diode laser for photoporation",

abstract = "The introduction and subsequent expression of foreign DNA inside living mammalian cells (transfection) is achieved by photoporation with a violet diode laser. We direct a compact 405 nm laser diode source into an inverted optical microscope configuration and expose cells to 0.3 mW for 40 ms. The localized optical power density of ~1200 MW/m2 is six orders of magnitude lower than that used in femtosecond photoporation (~104 TW/m2). The tightly focused laser beam (with a localised focal volume of ~10-19 m3 ) perforates the cell plasma membrane to allow uptake of plasmid DNA containing an antibiotic resistant gene as well as the green fluorescent protein (GFP) gene. Successfully transfected cells then expand into clonal groups which are used to create stable cell lines. The use of the violet diode laser offers a new and simple poration technique compatible with standard microscopes and is the simplest method of laser-assisted cell poration reported to date",

author = "Ben Agate and Lynn Paterson and Lake, {Tanya K.} and Morris, {J. E.} and Carruthers, {A. E.} and Brown, {Christian T. A.} and Wilson Sibbett and Kishan Dholakia and Muriel Comrie and R. Ferguson and Bryant, {P. E.} and Riches, {Andrew C.} and Gunn-Moore, {Frank J.}",

year = "2005",

doi = "10.1109/CLEOE.2005.1568410",

language = "English",

isbn = "0780389743",

pages = "634",

booktitle = "2005 Conference on Lasers and Electro-Optics Europe",

publisher = "OSA Publishing",

address = "United States",

note = "2005 Conference on Lasers and Elctro-Optics Europe ; Conference date: 12-06-2005 Through 17-06-2005",

}

Agate, B, Paterson, L, Lake, TK, Morris, JE, Carruthers, AE, Brown, CTA, Sibbett, W, Dholakia, K, Comrie, M, Ferguson, R, Bryant, PE, Riches, AC & Gunn-Moore, FJ 2005, Transfection of cells using a violet diode laser for photoporation. in 2005 Conference on Lasers and Electro-Optics Europe. OSA Publishing, pp. 634, 2005 Conference on Lasers and Elctro-Optics Europe, Munich, Germany, 12/06/05. https://doi.org/10.1109/CLEOE.2005.1568410

TY - GEN

T1 - Transfection of cells using a violet diode laser for photoporation

AU - Agate, Ben

AU - Paterson, Lynn

AU - Lake, Tanya K.

AU - Morris, J. E.

AU - Carruthers, A. E.

AU - Brown, Christian T. A.

AU - Sibbett, Wilson

AU - Dholakia, Kishan

AU - Comrie, Muriel

AU - Ferguson, R.

AU - Bryant, P. E.

AU - Riches, Andrew C.

AU - Gunn-Moore, Frank J.

PY - 2005

Y1 - 2005

N2 - The introduction and subsequent expression of foreign DNA inside living mammalian cells (transfection) is achieved by photoporation with a violet diode laser. We direct a compact 405 nm laser diode source into an inverted optical microscope configuration and expose cells to 0.3 mW for 40 ms. The localized optical power density of ~1200 MW/m2 is six orders of magnitude lower than that used in femtosecond photoporation (~104 TW/m2). The tightly focused laser beam (with a localised focal volume of ~10-19 m3 ) perforates the cell plasma membrane to allow uptake of plasmid DNA containing an antibiotic resistant gene as well as the green fluorescent protein (GFP) gene. Successfully transfected cells then expand into clonal groups which are used to create stable cell lines. The use of the violet diode laser offers a new and simple poration technique compatible with standard microscopes and is the simplest method of laser-assisted cell poration reported to date

AB - The introduction and subsequent expression of foreign DNA inside living mammalian cells (transfection) is achieved by photoporation with a violet diode laser. We direct a compact 405 nm laser diode source into an inverted optical microscope configuration and expose cells to 0.3 mW for 40 ms. The localized optical power density of ~1200 MW/m2 is six orders of magnitude lower than that used in femtosecond photoporation (~104 TW/m2). The tightly focused laser beam (with a localised focal volume of ~10-19 m3 ) perforates the cell plasma membrane to allow uptake of plasmid DNA containing an antibiotic resistant gene as well as the green fluorescent protein (GFP) gene. Successfully transfected cells then expand into clonal groups which are used to create stable cell lines. The use of the violet diode laser offers a new and simple poration technique compatible with standard microscopes and is the simplest method of laser-assisted cell poration reported to date

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U2 - 10.1109/CLEOE.2005.1568410

DO - 10.1109/CLEOE.2005.1568410

M3 - Conference contribution

AN - SCOPUS:42749101315

SN - 0780389743

SN - 9780780389748

SP - 634

BT - 2005 Conference on Lasers and Electro-Optics Europe

PB - OSA Publishing

T2 - 2005 Conference on Lasers and Elctro-Optics Europe

Y2 - 12 June 2005 through 17 June 2005

ER -

Transfection of cells using a violet diode laser for photoporation

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Conference

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