3D localized photoactivation of pa-GFP in living cells using two-photon interactions

Alberto Diaspro, Ilaria Testa, Mario Faretta, Raffaella Magrassi, Sara Barozzi, Dario Parazzoli, Giuseppe Vicidomini

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

We report about two-photon activation of 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. The aim in this work was to evaluate the use of two-photon interactions for confining the molecular switching of pa-GFP in the bright state. 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 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 and two-photon mode demonstrates clearly the better three-dimensional (3D) confinement and the possibility of selection of cell volumes of interest. This enables molecular trafficking studies at high signal to noise ratio.

Original languageEnglish
Title of host publicationAnnual International Conference of the IEEE Engineering in Medicine and Biology - Proceedings
Pages389-391
Number of pages3
DOIs
Publication statusPublished - 2006
Event28th Annual International Conference of the IEEE Engineering in Medicine and Biology Society, EMBS'06 - New York, NY, United States
Duration: Aug 30 2006Sep 3 2006

Other

Other28th Annual International Conference of the IEEE Engineering in Medicine and Biology Society, EMBS'06
CountryUnited States
CityNew York, NY
Period8/30/069/3/06

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ASJC Scopus subject areas

  • Bioengineering

Cite this

Diaspro, A., Testa, I., Faretta, M., Magrassi, R., Barozzi, S., Parazzoli, D., & Vicidomini, G. (2006). 3D localized photoactivation of pa-GFP in living cells using two-photon interactions. In Annual International Conference of the IEEE Engineering in Medicine and Biology - Proceedings (pp. 389-391). [4030035] https://doi.org/10.1109/IEMBS.2006.259967