Two-color multiphoton in vivo imaging with a femtosecond diamond Raman laser

作     者：Evan P Perillo Jeremy W Jarrett Yen-Liang Liu Ahmed Hassan Daniel C Fernée John R Goldak Andrei Bonteanu David J Spence Hsin-Chih Yeh Andrew K Dunn 

作者机构：Department of Biomedical EngineeringThe University of Texas at AustinTX 78712USA Department of PhysicsThe University of Texas at AustinTX 78712USA MQ PhotonicsDepartment of Physics and AstronomyMacquarie UniversitySydneyNSW 2109Australia 

出 版 物：《Light(Science & Applications)》 (光（科学与应用)（英文版）)

年 卷 期：2017年第6卷第1期

页      面：400-407页

核心收录：

学科分类：080901[工学-物理电子学] 0809[工学-电子科学与技术（可授工学、理学学位）] 08[工学] 080401[工学-精密仪器及机械] 0804[工学-仪器科学与技术] 0803[工学-光学工程] 

基　　金：supported by the National Institutes of Health(NS082518,CA193038,NS078791 and EB011556) the American Heart Association(14EIA18970041) Cancer Prevention Research Institute of Texas(RR160005) 

主　　题：microscopy nonlinear processes three-dimensional imaging ultrafast lasers 

摘      要：Two-color multiphoton microscopy through wavelength mixing of synchronized lasers has been shown to increase the spectral window of excitable fluorophores without the need for wavelength ***,most currently available dual output laser sources rely on the costly and complicated optical parametric generation *** this report,we detail a relatively simple and low cost diamond Raman laser pumped by a ytterbium fiber amplifier emitting at 1055 nm,which generates a first Stokes emission centered at 1240 nm with a pulse width of 100 *** two excitation wavelengths of 1055 and 1240 nm,along with the effective two-color excitation wavelength of 1140 nm,provide an almost complete coverage of fluorophores excitable within the range of 1000–1300 *** compared with 1055 nm excitation,two-color excitation at 1140 nm offers a 90%increase in signal for many far-red emitting fluorescent proteins(for example,tdKatushka2).We demonstrate multicolor imaging of tdKatushka2 and Hoechst 33342 via simultaneous two-color two-photon,and two-color three-photon microscopy in engineered 3D multicellular *** further discuss potential benefits and applications for two-color three-photon *** addition,we show that this laser system is capable of in vivo imaging in mouse cortex to nearly 1 mm in depth with two-color excitation.