Three-color femtosecond source for simultaneous excitation of three fluorescent proteins in two-photon fluorescence microscopy.

Content Provider	Europe PMC
Author	Wang, Ke Liu, Tzu-Ming Wu, Juwell Horton, Nicholas G. Lin, Charles P. Xu, Chris
Copyright Year	2012
Abstract	We demonstrate a fiber-based, three-color femtosecond source for simultaneous imaging of three fluorescent proteins (FPs) using two-photon fluorescence microscopy (2PM). The three excitation wavelengths at 775 nm, 864 nm and 950 nm, are obtained through second harmonic generation (SHG) of the 1550-nm pump laser and the 1728-nm and 1900-nm solitons generated through soliton self-frequency shift (SSFS) in a large-mode-area (LMA) fiber. These energetic pulses are well matched to the two-photon excitation peaks of red, cyan and yellow fluorescent proteins (TagRFPs, TagCFPs, and TagYFPs) for efficient excitation. We demonstrate simultaneous 2PM of human melanoma cells expressing a “rainbow” combination of these three fluorescent proteins.
Journal	Biomedical Optics Express
Volume Number	3
PubMed Central reference number	PMC3447541
Issue Number	9
PubMed reference number	23024893
e-ISSN	21567085
DOI	10.1364/BOE.3.001972
Language	English
Publisher	Optical Society of America
Publisher Date	2012-07-31
Access Restriction	Open
Rights License	This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 Unported License, which permits download and redistribution, provided that the original work is properly cited. This license restricts the article from being modified or used commercially. ©2012 Optical Society of America
Subject Keyword	(060.4370) Nonlinear optics, fibers (180.2520) Fluorescence microscopy (180.4315) Nonlinear microscopy (190.2620) Harmonic generation and mixing
Content Type	Text
Resource Type	Article
Subject	Biotechnology Atomic and Molecular Physics, and Optics