Three-photon excitation in fluorescence microscopy.
Three-photon excitation in fluorescence microscopy.
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DOI：
10.1117/12.229062
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发表时间：
1996-01-01
影响因子：
3.5
通讯作者：
Lakowicz, J R
中科院分区：
文献类型：
--
作者：
Hell, S W;Bahlmann, K;Lakowicz, J R
我们展示了实验证明的可行性扫描荧光显微镜的三光子激发。显示了用荧光团2，5-双（4-联苯基）恶唑（BBO）染色的聚苯乙烯珠的三光子激发荧光轴向图像。三光子激发在900 nm的激发波长下进行，并具有由锁模钛蓝宝石激光器提供的130 fs持续时间的脉冲。在350和450 nm之间收集荧光。荧光图像信号具有对激发功率的三阶依赖性，也提供了固有的3-D成像。一个三光子激发显微镜的分辨率增加了一个可比的双光子激发显微镜。
We show experiments proving the feasibility of scanning fluorescence microscopy by three-photon excitation. Three-photon excitation fluorescence axial images are shown of polystyrene beads stained with the fluorophore 2,5-bis(4-biphenyl)oxazole (BBO). Three-photon excitation is performed at an excitation wavelength of 900 nm and with pulses of 130 fs duration provided by a mode-locked titanium sapphire laser. Fluorescence is collected between 350 and 450 nm. The fluorescence image signal features a third-order dependence on the excitation power, also providing intrinsic 3-D imaging. The resolution of a three-photon excitation microscope is increased over that of a comparable two-photon excitation microscope.