Lee Sungon, Vinegoni Claudio, Sebas Matthew, Weissleder Ralph
1] Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, Richard B. Simches Research Center, 185 Cambridge Street, Boston 02114, USA [2] Interaction and Robotics Research Center, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul, Korea [3].
1] Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, Richard B. Simches Research Center, 185 Cambridge Street, Boston 02114, USA [2].
Sci Rep. 2014 Mar 28;4:4507. doi: 10.1038/srep04507.
Intravital fluorescence microscopy, through extended penetration depth and imaging resolution, provides the ability to image at cellular and subcellular resolution in live animals, presenting an opportunity for new insights into in vivo biology. Unfortunately, physiological induced motion components due to respiration and cardiac activity are major sources of image artifacts and impose severe limitations on the effective imaging resolution that can be ultimately achieved in vivo. Here we present a novel imaging methodology capable of automatically removing motion artifacts during intravital microscopy imaging of organs and orthotopic tumors. The method is universally applicable to different laser scanning modalities including confocal and multiphoton microscopy, and offers artifact free reconstructions independent of the physiological motion source and imaged organ. The methodology, which is based on raw data acquisition followed by image processing, is here demonstrated for both cardiac and respiratory motion compensation in mice heart, kidney, liver, pancreas and dorsal window chamber.
活体荧光显微镜通过扩展穿透深度和成像分辨率，能够在活体动物中以细胞和亚细胞分辨率进行成像，为深入了解体内生物学提供了契机。不幸的是，由于呼吸和心脏活动引起的生理诱导运动成分是图像伪影的主要来源，对最终能够在体内实现的有效成像分辨率施加了严重限制。在此，我们提出了一种新颖的成像方法，能够在对器官和原位肿瘤进行活体显微镜成像期间自动去除运动伪影。该方法普遍适用于包括共聚焦和多光子显微镜在内的不同激光扫描模式，并提供与生理运动源和成像器官无关的无伪影重建。这种基于原始数据采集然后进行图像处理的方法，在此展示了对小鼠心脏、肾脏、肝脏、胰腺和背窗室中的心脏和呼吸运动补偿。
