Active focus locking in an optically sectioning microscope utilizing a deformable membrane mirror
Active focus locking in an optically sectioning microscope utilizing a deformable membrane mirror
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DOI：
10.1364/ol.33.000419
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发表时间：
2008-03-01
期刊：
影响因子：
3.6
通讯作者：
Girkin, J. M.
中科院分区：
文献类型：
--
作者：
Poland, S. P.;Wright, A. J.;Girkin, J. M.
体内成像的一个重大挑战是去除运动伪影。这些运动（通常是由于呼吸和心脏相关的运动或表面化学反应）通常限于轴向方向，因此特征移入和移出焦平面。这为高分辨率光学切片成像技术如共焦和多光子显微术提出了一个真实的问题。为了克服这一点，我们已经开发了一个主动锁定的焦点跟踪系统的基础上周围的变形膜镜。这具有显着的优势，超过更传统的焦点跟踪技术，其中的显微镜物镜抖动，因为有源元件是不直接，或间接，与样品接触。为了检查操作限制并证明这种形式的焦点锁定的可能应用，针对两种不同的生物应用模拟样品振荡和移动。我们能够在400 μ m的范围内跟踪焦点（受压电安装物镜的范围限制），其焦点深度为0.31 μ m +/- 0.05 μ m。(C)2008年美国光学学会。
A significant challenge for in vivo imaging is to remove movement artifacts. These movements (typically due to either respiration and cardiac-related movement or surface chemical response) are normally limited to the axial direction, and hence features move in and out of the focal plane. This presents a real problem for high-resolution optically sectioned imaging techniques such as confocal and multiphoton microscopy. To overcome this we have developed an actively locked focus-tracking system based around a deformable membrane mirror. This has a significant advantage over more conventional focus-tracking techniques where the microscope objective is dithered, since the active element is not in direct, or indirect, contact with the sample. To examine the operational limits and to demonstrate possible applications for this form of focus locking, sample oscillation and movement are simulated for two different biological applications. We were able to track focus over a 400 mu m range (limited by the range of the piezomounted objective) with a rms precision on the focal depth of 0.31 mu m +/- 0.05 mu m. (C) 2008 Optical Society of America.