Chronic in vivo imaging in the mouse spinal cord using an implanted chamber.

Chronic in vivo imaging in the mouse spinal cord using an implanted chamber.
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DOI:
10.1038/nmeth.1856
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发表时间:
2012-01-22
期刊:
影响因子:
48
通讯作者:
Schaffer, Chris B.
Schaffer, Chris B.
中科院分区:
生物学1区
文献类型:
--
作者:
Farrar, Matthew J.;Bernstein, Ida M.;Schlafer, Donald H.;Cleland, Thomas A.;Fetcho, Joseph R.;Schaffer, Chris B.

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脊髓病理学的理解和治疗在一定程度上受到细胞水平上缺乏纵向体内成像策略的限制。我们开发了一种长期植入的脊髓腔和手术程序,适用于小鼠脊髓的延时体内多光子显微镜检查,而无需重复手术。术后5周以上常规重复成像,最多10次单独成像。我们既没有观察到运动功能缺陷,也没有神经病理学的脊髓作为一个结果的腔植入。使用该室,我们量化了小胶质细胞和传入轴突动力学激光诱导的脊髓损伤后,观察到大量的小胶质细胞浸润沿着一天内与一个异质的轴突残端枯死。通过在从几分钟到几个月的时间尺度上进行慢性成像研究,我们的方法为理解细胞动力学对损伤和治疗干预的反应提供了一个理想的平台。
Understanding and treatment of spinal cord pathology is limited in part by a lack of longitudinal in vivo imaging strategies at the cellular level. We developed a chronically implanted spinal chamber and surgical procedure suitable for time-lapse in vivo multiphoton microscopy of mouse spinal cord without the need for repeat surgical procedures. Repeated imaging was routinely achieved for more than five weeks post-operatively with up to ten separate imaging sessions. We observed neither motor function deficit nor neuropathology in the spinal cord as a result of chamber implantation. Using this chamber we quantified microglia and afferent axon dynamics following a laser-induced spinal cord lesion and observed massive microglia infiltration within one day along with a heterogeneous dieback of axon stumps. By enabling chronic imaging studies over timescales ranging from minutes to months, our method offers an ideal platform for understanding cellular dynamics in response to injury and therapeutic interventions.
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