Abstract
We report about a photoactivatable derivative of the Aequorea Victoria green fluorescent protein (paGFP). This special form of the molecule increases its fluorescence intensity when excited by 488 nm after irradiation with high intensity light at 413 nm 1. The aim in this work was to evaluate the use of two-photon interactions for activation of the molecules 2. Therefore experiments were performed using fixed and living cells which were expressing the paGFP fluorophore and microspheres whose surface was modified by specific adsorption of the chromophores. The latter objects were used to investigate the ability of different wavelengths to activate the paGFP due to the anticipated more homogeneous density distribution. The molecular switches were activated in a range of wavelength from 720 nm to 840 nm. The optimal wavelength for activation was then chosen for cell imaging. A comparison between the conventional activation with a single photon at 413 nm and two-photons demonstrates clearly the advantages using non linear processes: much smaller volume in the cell can be activated unlike to a whole cell activation in single photon excitation regime.
| Original language | English |
|---|---|
| Title of host publication | Proceedings of SPIE - The International Society for Optical Engineering |
| Volume | 6089 |
| DOIs | |
| Publication status | Published - 2006 |
| Event | Multiphoton Microscopy in the Biomedical Sciences VI - San Jose, CA, United States Duration: Jan 22 2006 → Jan 24 2006 |
Other
| Other | Multiphoton Microscopy in the Biomedical Sciences VI |
|---|---|
| Country | United States |
| City | San Jose, CA |
| Period | 1/22/06 → 1/24/06 |
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Keywords
- Fluorescence microscopy
- PA-GFP
- Two-photon activation
- Two-photon excitation
ASJC Scopus subject areas
- Electrical and Electronic Engineering
- Condensed Matter Physics
Cite this
T2P-GFP : Two-photon photo-activation of PA-GFP in the 720-840 nm spectral region. / Testa, Ilaria; Schneider, Marc; Barozzi, Sara; Vicidomini, Giuseppe; Parazzoli, Dario; Faretta, Mario; Diaspro, Alberto.
Proceedings of SPIE - The International Society for Optical Engineering. Vol. 6089 2006. 608912.Research output: Chapter in Book/Report/Conference proceeding › Conference contribution
}
TY - GEN
T1 - T2P-GFP
T2 - Two-photon photo-activation of PA-GFP in the 720-840 nm spectral region
AU - Testa, Ilaria
AU - Schneider, Marc
AU - Barozzi, Sara
AU - Vicidomini, Giuseppe
AU - Parazzoli, Dario
AU - Faretta, Mario
AU - Diaspro, Alberto
PY - 2006
Y1 - 2006
N2 - We report about a photoactivatable derivative of the Aequorea Victoria green fluorescent protein (paGFP). This special form of the molecule increases its fluorescence intensity when excited by 488 nm after irradiation with high intensity light at 413 nm 1. The aim in this work was to evaluate the use of two-photon interactions for activation of the molecules 2. Therefore experiments were performed using fixed and living cells which were expressing the paGFP fluorophore and microspheres whose surface was modified by specific adsorption of the chromophores. The latter objects were used to investigate the ability of different wavelengths to activate the paGFP due to the anticipated more homogeneous density distribution. The molecular switches were activated in a range of wavelength from 720 nm to 840 nm. The optimal wavelength for activation was then chosen for cell imaging. A comparison between the conventional activation with a single photon at 413 nm and two-photons demonstrates clearly the advantages using non linear processes: much smaller volume in the cell can be activated unlike to a whole cell activation in single photon excitation regime.
AB - We report about a photoactivatable derivative of the Aequorea Victoria green fluorescent protein (paGFP). This special form of the molecule increases its fluorescence intensity when excited by 488 nm after irradiation with high intensity light at 413 nm 1. The aim in this work was to evaluate the use of two-photon interactions for activation of the molecules 2. Therefore experiments were performed using fixed and living cells which were expressing the paGFP fluorophore and microspheres whose surface was modified by specific adsorption of the chromophores. The latter objects were used to investigate the ability of different wavelengths to activate the paGFP due to the anticipated more homogeneous density distribution. The molecular switches were activated in a range of wavelength from 720 nm to 840 nm. The optimal wavelength for activation was then chosen for cell imaging. A comparison between the conventional activation with a single photon at 413 nm and two-photons demonstrates clearly the advantages using non linear processes: much smaller volume in the cell can be activated unlike to a whole cell activation in single photon excitation regime.
KW - Fluorescence microscopy
KW - PA-GFP
KW - Two-photon activation
KW - Two-photon excitation
UR - http://www.scopus.com/inward/record.url?scp=33646168489&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33646168489&partnerID=8YFLogxK
U2 - 10.1117/12.661104
DO - 10.1117/12.661104
M3 - Conference contribution
AN - SCOPUS:33646168489
SN - 0819461318
SN - 9780819461315
VL - 6089
BT - Proceedings of SPIE - The International Society for Optical Engineering
ER -