Department of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia, Genova, Italy.
Opt Lett. 2011 Sep 1;36(17):3503-5. doi: 10.1364/OL.36.003503.
The acquisition of high-resolution images in three dimensions is of utmost importance for the morphological and functional investigation of biological tissues. Here, we present a laser scanning two-photon microscope with remote and motionless control of the focus position. The movement of the excitation spot along the propagation direction is achieved by shaping the laser wavefront with a spatial light modulator. Depending on the optical properties of the objective in use, this approach allows z movements in a range of tens to hundreds of micrometers with small changes of the point spread function. We applied this technique for the three-dimensional (3D) imaging of fluorescent cells in the mouse neocortex in vivo. The presented system bypasses the limitations of microscopes based on moving objectives, enabling high-resolution inertia-free 3D imaging.
获取高分辨率的三维图像对于生物组织的形态和功能研究至关重要。在这里，我们展示了一种具有远程和无移动控制焦点位置的激光扫描双光子显微镜。通过使用空间光调制器对激光波前进行整形，可以实现激发光斑沿传播方向的移动。根据所使用物镜的光学特性，这种方法可以在数十到数百微米的范围内进行 z 移动，并且点扩散函数的变化很小。我们将该技术应用于活体小鼠新皮层中荧光细胞的三维（3D）成像。所提出的系统克服了基于移动物镜的显微镜的局限性，实现了高分辨率无惯性的 3D 成像。
