Subcellular Motion Compensation for Minimally Invasive Microscopy, In Vivo Evidence for Oxygen Gradients in Resting Muscle
Subcellular Motion Compensation for Minimally Invasive Microscopy, In Vivo Evidence for Oxygen Gradients in Resting Muscle
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DOI：
10.1161/circresaha.109.211946
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发表时间：
2010-04-02
影响因子：
20.1
通讯作者：
Balaban, Robert S.
中科院分区：
文献类型：
--
作者：
Schroeder, James L.;Luger-Hamer, Merav;Balaban, Robert S.
理由：体内显微镜旨在观察生理相关背景下的动态亚细胞过程。体内光学显微镜的一个主要限制是组织运动，它阻碍了生理时间过程观察或图像平均。 目的：开发和演示运动补偿方法，可以自动跟踪生物组织内的图像平面，包括与血流大幅变化相关的组织位移，并评估全局缺氧对线粒体局部动力学和稳态水平的影响 NAD(P)H.方法和结果：使用具有实时前瞻性跟踪和回顾性图像处理功能的动态光学显微镜，通过细胞对各种扰动的反应来收集高分辨率图像。在体内检查了亚细胞对缺氧的代谢反应。最靠近毛细血管的线粒体在休息时 (67 +/- 3%) 比原纤维内线粒体 (83 +/- 3%; P
Rationale: In vivo microscopy seeks to observe dynamic subcellular processes in a physiologically relevant context. A primary limitation of optical microscopy in vivo is tissue motion, which prevents physiological time course observations or image averaging.Objective: To develop and demonstrate motion compensation methods that can automatically track image planes within biological tissues, including the tissue displacements associated with large changes in blood flow, and to evaluate the effect of global hypoxia on the regional kinetics and steady state levels of mitochondrial NAD(P)H.Methods and Results: A dynamic optical microscope, with real-time prospective tracking and retrospective image processing, was used collect high-resolution images through cellular responses to various perturbations. The subcellular metabolic response to hypoxia was examined in vivo. Mitochondria closest to the capillaries were significantly more oxidized at rest (67 +/- 3%) than the intrafibrillar mitochondria (83 +/- 3%; P