title: Multifarious control of two-photon excitation of multiple fluorophores achieved by phase modulation of ultra-broadband laser pulses.
abstract: We propose two-photon excited fluorescence (TPEF) microscopy employing a novel phase modulation technique of ultra-broadband laser pulses, which allows the relative excitation of an individual fluorophore with respect to other fluorophores. This technique is based on the generation of multi-wavelength pulse train, which independently interacts with each fluorophore. Our technique is applied to dual-color imaging of cells expressing two types of fluorescent proteins. We achieve the selective excitation of one over the other and vice versa. The product of the maximum contrast ratios exceeds 100. We also demonstrate yielded equal excitation rates in the simultaneous excitation. By the selective excitation of a donor fluorescent protein, fluorescence resonance energy transfer imaging is also achieved.
authors: Laser Technology Laboratory, RIKEN Advanced Science Institute, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan. kisobe@riken.jp
authors: Keisuke
authors: K
authors: Isobe
authors: Akira
authors: A
authors: Suda
authors: Masahiro
authors: M
authors: Tanaka
authors: Fumihiko
authors: F
authors: Kannari
authors: Hiroyuki
authors: H
authors: Kawano
authors: Hideaki
authors: H
authors: Mizuno
authors: Atsushi
authors: A
authors: Miyawaki
authors: Katsumi
authors: K
authors: Midorikawa
mesh_terms: D003198:Computer Simulation; D017076:Computer-Aided Design; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007834:Lasers; D008027:Light; D008029:Lighting / Q000295:instrumentation*; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation* / Q000379:methods; D008962:Models, Theoretical; D055096:Optical Devices; D012542:Scattering, Radiation