Automated filtering of intrinsic movement artifacts during two-photon intravital microscopy.
Automated filtering of intrinsic movement artifacts during two-photon intravital microscopy.
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DOI：
10.1371/journal.pone.0053942
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Lacroix S
中科院分区：
文献类型：
--
作者：
Soulet D;Paré A;Coste J;Lacroix S
使用双光子显微镜的体内成像是在短时间或长时间内探索生物组织深处的生理事件动态的重要工具。该技术提供的新功能（例如高组织渗透率，低毒性）开启了现代生物医学研究的全新时代。然而，在活体动物组织中使用这种有前途的技术的潜力受到由心脏和呼吸周期以及肌肉和血管张力引起的固有不规则运动的极大限制。在这里，我们使用一种名为Intravital_Microscopy_Mapping的新自动化程序显示了活体小鼠的大脑，脊髓，坐骨神经和肌间神经丛的实时成像，该程序可以从延时视频中删除因运动伪影而损坏的帧。我们的方法涉及在特定参考通道中生成针对预先计算的参考帧的相异性分数，从而允许对失真、失焦或平移帧进行门控。由于该算法检测不规则动物运动引起的图像失真的不均匀峰值，因此宏允许对图像序列进行快速有效的滤波。此外，在宏中还实现了额外的功能，例如XY配准、通道减法、具有最大强度投影的扩展视野、具有平均强度投影的降噪以及自动时间戳和比例尺叠加。因此，ImageJ的Intravital_Microscopy_Microscopy宏为在易于出现运动伪影的组织中执行体内双光子成像的生物学家提供了方便的工具。
In vivo imaging using two-photon microscopy is an essential tool to explore the dynamic of physiological events deep within biological tissues for short or extended periods of time. The new capabilities offered by this technology (e.g. high tissue penetrance, low toxicity) have opened a whole new era of investigations in modern biomedical research. However, the potential of using this promising technique in tissues of living animals is greatly limited by the intrinsic irregular movements that are caused by cardiac and respiratory cycles and muscular and vascular tone. Here, we show real-time imaging of the brain, spinal cord, sciatic nerve and myenteric plexus of living mice using a new automated program, named Intravital_Microscopy_Toolbox, that removes frames corrupted with motion artifacts from time-lapse videos. Our approach involves generating a dissimilarity score against precalculated reference frames in a specific reference channel, thus allowing the gating of distorted, out-of-focus or translated frames. Since the algorithm detects the uneven peaks of image distortion caused by irregular animal movements, the macro allows a fast and efficient filtering of the image sequence. In addition, extra features have been implemented in the macro, such as XY registration, channel subtraction, extended field of view with maximum intensity projection, noise reduction with average intensity projections, and automated timestamp and scale bar overlay. Thus, the Intravital_Microscopy_Toolbox macro for ImageJ provides convenient tools for biologists who are performing in vivo two-photon imaging in tissues prone to motion artifacts.
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影响因子：
64.5
作者：
Pittet MJ;Weissleder R
通讯作者：
Weissleder R
影响因子：
56.9
作者：
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通讯作者：
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影响因子：
3
作者：
Hecker, David;Kappler, Joachim;Alt, Wolfgang
通讯作者：
Alt, Wolfgang