University of British Columbia, Vancouver, British Columbia, Canada.
Stroke. 2010 Oct;41(10 Suppl):S117-23. doi: 10.1161/STROKEAHA.110.594648.
The recent application of 2-photon microscopy to biological specimens has allowed investigators to examine individual synapses within live animals. The gain in resolution over conventional in vivo imaging techniques has been several orders of magnitude. We outline steps for the preparation and maintenance of animals for 2-photon microscopy of fine brain structure. We discuss the in vivo resolution of the method and the ability to image blood flow and synaptic structure in vivo. Applications of in vivo 2-photon microscopy include the study of synapse turnover in adult animals under normal conditions and during pathology such as stroke. In the case of stroke, 2-photon imaging has revealed marked swelling of dendrites and loss of spines within minutes of ischemic onset. Surprisingly, restoration of blood flow during reperfusion was associated with a return of relatively normal structure. Over longer time scales, 2-photon imaging revealed elevated rates of synaptogenesis within peri-infarct tissues recovering from stroke. These results provide an example of how high-resolution in vivo microscopy can be used to provide insight into both the acute pathology and recovery from stroke damage.
最近将双光子显微镜应用于生物样本，使得研究人员能够在活体动物中检查单个突触。与传统的活体成像技术相比，分辨率提高了几个数量级。我们概述了准备和维持动物用于精细脑结构双光子显微镜的步骤。我们讨论了该方法的活体分辨率以及在活体中成像血流和突触结构的能力。活体双光子显微镜的应用包括在正常条件下和在病理如中风期间研究成年动物中的突触周转。在中风的情况下，双光子成像揭示了在缺血发作后几分钟内树突和棘突明显肿胀和丢失。令人惊讶的是，在再灌注期间恢复血流与相对正常结构的恢复相关联。在更长的时间尺度上，双光子成像显示出在从中风中恢复的梗塞周围组织中突触发生的速率升高。这些结果提供了一个例子，说明如何使用高分辨率活体显微镜来深入了解中风损伤的急性病理和恢复。
