Multiphoton in vivo imaging with a femtosecond semiconductor disk laser.
Multiphoton in vivo imaging with a femtosecond semiconductor disk laser.
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DOI：
10.1364/boe.8.003213
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发表时间：
2017-07
影响因子：
3.4
通讯作者：
F. Voigt;F. Emaury;P. Bethge;D. Waldburger;S. Link;Stefano Carta;Alexander van der Bourg;F. Helmchen;U. Keller
中科院分区：
文献类型：
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作者：
F. Voigt;F. Emaury;P. Bethge;D. Waldburger;S. Link;Stefano Carta;Alexander van der Bourg;F. Helmchen;U. Keller
我们使用超快二极管泵浦的半导体圆盘激光器（SDL）来演示多光子显微镜中的几个应用。超快SDL是基于一个光泵垂直外腔面发射激光器（VECSEL）被动锁模与半导体可饱和吸收镜（SESAM），并产生170 fs的脉冲在1027 nm的中心波长与1.63 GHz的重复率。我们证明了这种激光器的结构和功能的多光子在体内成像在果蝇幼虫和小鼠的各种荧光团（包括mKate 2，tdTomato，得克萨斯红，OGB-1，和R-CaMP 1.07）和内源性二次谐波产生在肌细胞肌节的适用性。当我们将平均功率增加到理论预测的4.5倍时，我们可以证明与标准80 MHz Ti：Sapphire激光器相比具有等效的信号水平。此外，我们比较了固定果蝇幼虫的两种激光系统的漂白性能，发现尽管脉冲重复率差异很大，但漂白动力学相似。我们的研究结果突出了超快二极管泵浦的SDLs的巨大潜力，创造一个具有成本效益和紧凑的替代光源相比，标准的钛：蓝宝石激光器的多光子成像。
We use an ultrafast diode-pumped semiconductor disk laser (SDL) to demonstrate several applications in multiphoton microscopy. The ultrafast SDL is based on an optically pumped Vertical External Cavity Surface Emitting Laser (VECSEL) passively mode-locked with a semiconductor saturable absorber mirror (SESAM) and generates 170-fs pulses at a center wavelength of 1027 nm with a repetition rate of 1.63 GHz. We demonstrate the suitability of this laser for structural and functional multiphoton in vivo imaging in both Drosophila larvae and mice for a variety of fluorophores (including mKate2, tdTomato, Texas Red, OGB-1, and R-CaMP1.07) and for endogenous second-harmonic generation in muscle cell sarcomeres. We can demonstrate equivalent signal levels compared to a standard 80-MHz Ti:Sapphire laser when we increase the average power by a factor of 4.5 as predicted by theory. In addition, we compare the bleaching properties of both laser systems in fixed Drosophila larvae and find similar bleaching kinetics despite the large difference in pulse repetition rates. Our results highlight the great potential of ultrafast diode-pumped SDLs for creating a cost-efficient and compact alternative light source compared to standard Ti:Sapphire lasers for multiphoton imaging.