Details
| Original language | English |
|---|---|
| Title of host publication | Optical Trapping and Optical Micromanipulation VII |
| Publication status | Published - 27 Aug 2010 |
| Event | Optical Trapping and Optical Micromanipulation VII - San Diego, CA, United States Duration: 1 Aug 2010 → 5 Aug 2010 |
Publication series
| Name | Proceedings of SPIE - The International Society for Optical Engineering |
|---|---|
| Volume | 7762 |
| ISSN (Print) | 0277-786X |
Abstract
The delivery of extra cellular molecules into cells is essential for cell manipulation. For this purpose genetic materials (DNA/RNA) or proteins have to overcome the impermeable cell membrane. To increase the delivery efficiency and cell viability of common methods different nano-and micro material based approaches were applied. To manipulate the cells, the membrane is in contact with the biocompatible material. Due to a field enhancement of the laser light at the material and the resulting effect the cell membrane gets perforated and extracellular molecules can diffuse into the cytoplasm. Membrane impermeable dyes, fluorescent labelled siRNA, as well as plasmid vectors encoded for GFP expression were used as an indicator for successful perforation or transfection, respectively. Dependent on the used material, perforation efficiencies over 90 % with a cell viability of about 80 % can be achieved. Additionally, we observed similar efficiencies for siRNA transfection. Due to the larger molecule size and the essential transport of the DNA into the nucleus cells are more difficult to transfect with GFP plasmid vectors. Proof of principle experiments show promising and adequate efficiencies by applying micro materials for plasmid vector transfection. For all methods a weakly focused fs laser beam is used to enable a high manipulation throughput for adherent and suspension cells. Furthermore, with these alternative optical manipulation methods it is possible to perforate the membrane of sensitive cell types such as primary and stem cells with a high viability.
Keywords
- Biophotonics, Cell manipulation, GFP, Membrane permeabilization, Nanomaterial, Nanoparticles, Perforation, Plasmonics, Transfection, Ultrashort laser pulses
ASJC Scopus subject areas
- Materials Science(all)
- Electronic, Optical and Magnetic Materials
- Physics and Astronomy(all)
- Condensed Matter Physics
- Computer Science(all)
- Computer Science Applications
- Mathematics(all)
- Applied Mathematics
- Engineering(all)
- Electrical and Electronic Engineering
Cite this
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- Harvard
- Apa
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- BibTeX
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Optical Trapping and Optical Micromanipulation VII. 2010. 77623G (Proceedings of SPIE - The International Society for Optical Engineering; Vol. 7762).
Research output: Chapter in book/report/conference proceeding › Conference contribution › Research › peer review
}
TY - GEN
T1 - Ultrashort laser pulse cell manipulation using nano-and micro-materials
AU - Schomaker, Markus
AU - Killian, Doreen
AU - Willenbrock, Saskia
AU - Diebold, Eric
AU - Mazur, Eric
AU - Bintig, Willem
AU - Ngezahayo, Anaclet
AU - Nolte, Ingo
AU - Murua Escobar, Hugo
AU - Junghanß, Christian
AU - Lubatschowski, Holger
AU - Heisterkamp, Alexander
PY - 2010/8/27
Y1 - 2010/8/27
N2 - The delivery of extra cellular molecules into cells is essential for cell manipulation. For this purpose genetic materials (DNA/RNA) or proteins have to overcome the impermeable cell membrane. To increase the delivery efficiency and cell viability of common methods different nano-and micro material based approaches were applied. To manipulate the cells, the membrane is in contact with the biocompatible material. Due to a field enhancement of the laser light at the material and the resulting effect the cell membrane gets perforated and extracellular molecules can diffuse into the cytoplasm. Membrane impermeable dyes, fluorescent labelled siRNA, as well as plasmid vectors encoded for GFP expression were used as an indicator for successful perforation or transfection, respectively. Dependent on the used material, perforation efficiencies over 90 % with a cell viability of about 80 % can be achieved. Additionally, we observed similar efficiencies for siRNA transfection. Due to the larger molecule size and the essential transport of the DNA into the nucleus cells are more difficult to transfect with GFP plasmid vectors. Proof of principle experiments show promising and adequate efficiencies by applying micro materials for plasmid vector transfection. For all methods a weakly focused fs laser beam is used to enable a high manipulation throughput for adherent and suspension cells. Furthermore, with these alternative optical manipulation methods it is possible to perforate the membrane of sensitive cell types such as primary and stem cells with a high viability.
AB - The delivery of extra cellular molecules into cells is essential for cell manipulation. For this purpose genetic materials (DNA/RNA) or proteins have to overcome the impermeable cell membrane. To increase the delivery efficiency and cell viability of common methods different nano-and micro material based approaches were applied. To manipulate the cells, the membrane is in contact with the biocompatible material. Due to a field enhancement of the laser light at the material and the resulting effect the cell membrane gets perforated and extracellular molecules can diffuse into the cytoplasm. Membrane impermeable dyes, fluorescent labelled siRNA, as well as plasmid vectors encoded for GFP expression were used as an indicator for successful perforation or transfection, respectively. Dependent on the used material, perforation efficiencies over 90 % with a cell viability of about 80 % can be achieved. Additionally, we observed similar efficiencies for siRNA transfection. Due to the larger molecule size and the essential transport of the DNA into the nucleus cells are more difficult to transfect with GFP plasmid vectors. Proof of principle experiments show promising and adequate efficiencies by applying micro materials for plasmid vector transfection. For all methods a weakly focused fs laser beam is used to enable a high manipulation throughput for adherent and suspension cells. Furthermore, with these alternative optical manipulation methods it is possible to perforate the membrane of sensitive cell types such as primary and stem cells with a high viability.
KW - Biophotonics
KW - Cell manipulation
KW - GFP
KW - Membrane permeabilization
KW - Nanomaterial
KW - Nanoparticles
KW - Perforation
KW - Plasmonics
KW - Transfection
KW - Ultrashort laser pulses
UR - http://www.scopus.com/inward/record.url?scp=77958149767&partnerID=8YFLogxK
U2 - 10.1117/12.869054
DO - 10.1117/12.869054
M3 - Conference contribution
AN - SCOPUS:77958149767
SN - 9780819482587
T3 - Proceedings of SPIE - The International Society for Optical Engineering
BT - Optical Trapping and Optical Micromanipulation VII
T2 - Optical Trapping and Optical Micromanipulation VII
Y2 - 1 August 2010 through 5 August 2010
ER -