Details
| Original language | English |
|---|---|
| Pages (from-to) | 2219-2229 |
| Number of pages | 11 |
| Journal | Optics Express |
| Volume | 18 |
| Issue number | 3 |
| Publication status | Published - 19 Jan 2010 |
Abstract
Fs-laser based opto-perforation is a gentle method for gene transfer into sensitive cells such as stem cells or primary cells. The high selectivity and the low damage to the cell lead to a high efficiency of transfection. However, there are side effects which induce stress to the cell due to the exchange of intra- and extracellular media as well as the disintegration of the structure of biomolecules resulting from the laser exposure. Moreover, the mechanisms of the optical transfection are still unclear. In this paper, we present our study on calcium (Ca2+) homeostasis during cell surgery, especially during laser induced membrane perforation. We show that the manipulation of cells can induce an increase in the cytosolic Ca2+ concentration. This increase was not observed if the manipulation of the cells was performed in absence of the extracellular calcium indicating the importance of the Ca2+ uptake. We found, that the uptake of extracellular Ca2+ strongly depends on the repetition rate and the irradiation time of the laser pulses. The exposure for several seconds to kHz pulses even induces Ca2+ induced Ca2+ release. Dependent on the location of perforation, probably in the vicinity of an intracellular Ca2+ stock, an instantaneous intracellular Ca2+ release can be induced. Since Ca2+ could be involved in negative side effect by cell surgery, we propose an application of the optoperforation technique in nominal Ca2+-free external solution.
ASJC Scopus subject areas
- Physics and Astronomy(all)
- Atomic and Molecular Physics, and Optics
Cite this
- Standard
- Harvard
- Apa
- Vancouver
- BibTeX
- RIS
In: Optics Express, Vol. 18, No. 3, 19.01.2010, p. 2219-2229.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Fs-laser-induced Ca2+ concentration change during membrane perforation for cell transfection
AU - Baumgart, Judith
AU - Bintig, Willem
AU - Ngezahayo, Anaclet
AU - Lubatschowski, Holger
AU - Heisterkamp, Alexander
PY - 2010/1/19
Y1 - 2010/1/19
N2 - Fs-laser based opto-perforation is a gentle method for gene transfer into sensitive cells such as stem cells or primary cells. The high selectivity and the low damage to the cell lead to a high efficiency of transfection. However, there are side effects which induce stress to the cell due to the exchange of intra- and extracellular media as well as the disintegration of the structure of biomolecules resulting from the laser exposure. Moreover, the mechanisms of the optical transfection are still unclear. In this paper, we present our study on calcium (Ca2+) homeostasis during cell surgery, especially during laser induced membrane perforation. We show that the manipulation of cells can induce an increase in the cytosolic Ca2+ concentration. This increase was not observed if the manipulation of the cells was performed in absence of the extracellular calcium indicating the importance of the Ca2+ uptake. We found, that the uptake of extracellular Ca2+ strongly depends on the repetition rate and the irradiation time of the laser pulses. The exposure for several seconds to kHz pulses even induces Ca2+ induced Ca2+ release. Dependent on the location of perforation, probably in the vicinity of an intracellular Ca2+ stock, an instantaneous intracellular Ca2+ release can be induced. Since Ca2+ could be involved in negative side effect by cell surgery, we propose an application of the optoperforation technique in nominal Ca2+-free external solution.
AB - Fs-laser based opto-perforation is a gentle method for gene transfer into sensitive cells such as stem cells or primary cells. The high selectivity and the low damage to the cell lead to a high efficiency of transfection. However, there are side effects which induce stress to the cell due to the exchange of intra- and extracellular media as well as the disintegration of the structure of biomolecules resulting from the laser exposure. Moreover, the mechanisms of the optical transfection are still unclear. In this paper, we present our study on calcium (Ca2+) homeostasis during cell surgery, especially during laser induced membrane perforation. We show that the manipulation of cells can induce an increase in the cytosolic Ca2+ concentration. This increase was not observed if the manipulation of the cells was performed in absence of the extracellular calcium indicating the importance of the Ca2+ uptake. We found, that the uptake of extracellular Ca2+ strongly depends on the repetition rate and the irradiation time of the laser pulses. The exposure for several seconds to kHz pulses even induces Ca2+ induced Ca2+ release. Dependent on the location of perforation, probably in the vicinity of an intracellular Ca2+ stock, an instantaneous intracellular Ca2+ release can be induced. Since Ca2+ could be involved in negative side effect by cell surgery, we propose an application of the optoperforation technique in nominal Ca2+-free external solution.
UR - http://www.scopus.com/inward/record.url?scp=76149089097&partnerID=8YFLogxK
U2 - 10.1364/OE.18.002219
DO - 10.1364/OE.18.002219
M3 - Article
C2 - 20174050
AN - SCOPUS:76149089097
VL - 18
SP - 2219
EP - 2229
JO - Optics Express
JF - Optics Express
SN - 1094-4087
IS - 3
ER -