Research Papers

Laser-assisted optoporation of single cells

[+] Author Affiliations
Herbert Schneckenburger

Institut fu¨r Angewandte Forschung, Fachhochschule Aalen, 73428?Aalen, Germany

and Universita¨t Ulm, Institut fu¨r Lasertechnologien in der Medizin und Messtechnik, Helmholzstrasse 12, 89081?Ulm, Germany

Anita Hendinger

Institut fu¨r Angewandte Forschung, Fachhochschule Aalen, 73428?Aalen, Germany

Reinhard Sailer, Wolfgang S. L. Strauss

Universita¨t Ulm, Institut fu¨r Lasertechnologien in der Medizin und Messtechnik, Helmholtzstrasse 12, 89081?Ulm, Germany

Michael Schmitt

Universita¨t Ulm, Medizinische Klinik, Innere Medizin III, Robert-Koch-Strasse 8, 89081?Ulm, Germany

J. Biomed. Opt. 7(3), 410-416 (Jul 01, 2002). doi:10.1117/1.1485758
History: Received Sep. 19, 2001; Revised Feb. 26, 2002; Accepted Mar. 1, 2002; Online August 01, 2002
Text Size: A A A

The plasma membrane of Chinese hamster ovary cells was made permeable using the focused beam of an argon ion laser (488 nm) and phenol red as a light absorbing dye. Small circular dark spots on the cell surface appeared immediately after laser irradiation and disappeared within about 5 min. They were related to transient changes in membrane properties, which could be visualized using the fluorescent marker laurdan, and were probably due to a local increase in temperature. According to a colony forming assay, cell viability was maintained by using light doses up to 2.5 MJ/cm2 applied for 1 s. In addition to measurements of the efflux of the cytoplasmic marker calcein, cell transfection using a green fluorescent protein (GFP) coding plasmid was studied: brightly fluorescent GFP with an emission maximum around 510 nm was observed within part of the cells after 24 h. The transfection rates after laser irradiation were around 30% for younger subcultures and less than 10% for aging cells. This may be due to age dependent changes in the phase transition of membrane lipids from gel phase to liquid crystalline phase. High transfection rates, visual control and universality towards various cell lines are possibly the main advantages of laser-assisted optoporation in comparison with presently existing methods of cell transfection. © 2002 Society of Photo-Optical Instrumentation Engineers.

© 2002 Society of Photo-Optical Instrumentation Engineers

Citation

Herbert Schneckenburger ; Anita Hendinger ; Reinhard Sailer ; Wolfgang S. L. Strauss and Michael Schmitt
"Laser-assisted optoporation of single cells", J. Biomed. Opt. 7(3), 410-416 (Jul 01, 2002). ; http://dx.doi.org/10.1117/1.1485758


Tables

Access This Article
Sign in or Create a personal account to Buy this article ($20 for members, $25 for non-members).

Some tools below are only available to our subscribers or users with an online account.

Related Content

Customize your page view by dragging & repositioning the boxes below.

Related Book Chapters

Topic Collections

PubMed Articles
Advertisement
  • Don't have an account?
  • Subscribe to the SPIE Digital Library
  • Create a FREE account to sign up for Digital Library content alerts and gain access to institutional subscriptions remotely.
Access This Article
Sign in or Create a personal account to Buy this article ($20 for members, $25 for non-members).
Access This Proceeding
Sign in or Create a personal account to Buy this article ($15 for members, $18 for non-members).
Access This Chapter

Access to SPIE eBooks is limited to subscribing institutions and is not available as part of a personal subscription. Print or electronic versions of individual SPIE books may be purchased via SPIE.org.