Bewersdorf J, Pick R, Hell S W
Opt Lett. 1998;23(9):655-7. doi: 10.1364/ol.23.000655.
We present a real-time, direct-view multiphoton excitation fluorescence microscope that provides three-dimensional imaging at high resolution. Using a rotating microlens disk, we split the near-infrared light of a mode-locked titanium-sapphire laser into an array of beams that are transformed into an array of high-aperture foci at the object. We typically scan at 225 frames per second and image the fluorescence with a camera that reads out the images at video rate. For 1.4 aperture oil and 1.2 water immersion lenses at 780-nm excitation we obtained axial resolutions of 0.84 and 1.4mum , respectively, which are similar to that of a single-beam two-photon microscope. Compared with the latter setup, our system represents a 40-100-fold increase in efficiency, or imaging speed. Moreover, it permits the observation with the eye of high-resolution two-photon images of (live) samples.
我们展示了一种实时、直视多光子激发荧光显微镜，它能提供高分辨率的三维成像。通过使用旋转微透镜盘，我们将锁模钛宝石激光器的近红外光分成一束束光束阵列，这些光束在物体处转化为高孔径焦点阵列。我们通常以每秒225帧的速度进行扫描，并用一台以视频速率读出图像的相机对荧光进行成像。对于在780纳米激发下的1.4孔径油浸物镜和1.2孔径水浸物镜，我们分别获得了0.84微米和1.4微米的轴向分辨率，这与单光束双光子显微镜的分辨率相似。与后一种设置相比，我们的系统效率提高了40至100倍，即成像速度提高了40至100倍。此外，它还允许用肉眼观察（活体）样品的高分辨率双光子图像。
