@inbook{7b8829dac1b64d98a37bd11a0a1e6880,
title = "Deep-tissue multi-photon fluorescence lifetime microscopy for intravital imaging of protein-protein interactions",
abstract = "Fluorescent lifetime imaging microscopy (FLIM) has proven to be a valuable tool in beating the Rayleigh criterion for light microscopy by measuring Forster resonance energy transfer (FRET) between two fluorophores. Applying multiphoton FLIM, we previously showed in a human breast cancer cell line that recycling of a membrane receptor-green fluorescent protein fusion is enhanced concomitantly with the formation of a receptor: protein kinase C alpha complex in the endosomal compartment. We have extended this established technique to probe direct protein-protein interactions also in vivo. Therefore, we used various expressible fluorescent tags fused to membrane receptor molecules in order to generate stable two-colour breast carcinoma cell lines via controlled retroviral infection. We used these cell lines for establishing a xenograft tumour model in immune-compromised Nude mice. Using this animal model in conjunction with scanning Ti:Sapphire laser-based two-photon excitation, we established deep-tissue multiphoton FLIM in vivo. For the first time, this novel technique enables us to directly assess donor fluorescence lifetime changes in vivo and we show the application of this method for intravital imaging of direct protein-protein interactions.",
author = "Fruhwirth, {G. O.} and Matthews, {D. R.} and A. Brock and M. Keppler and B. Vojnovic and T. Ng and S. Ameer-Beg",
year = "2009",
language = "English",
isbn = "978-0-8194-7429-2",
series = "MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES IX",
publisher = "SPIE - INT SOC OPTICAL ENGINEERING",
booktitle = "Unknown",
note = "Conference on Multiphoton Microscopy in the Biomedical Sciences IX ; Conference date: 25-01-2009 Through 27-01-2009",
}
