Department of Biological Sciences, Purdue University, 915 West State Street, West Lafayette, IN 47907-2054, USA.
J Neurosci Methods. 2011 Sep 30;201(1):67-77. doi: 10.1016/j.jneumeth.2011.07.012. Epub 2011 Jul 23.
Accurate assessment of brain-implantable microdevice bio-integration remains a formidable challenge. Prevailing histological methods require device extraction prior to tissue processing, often disrupting and removing the tissue of interest which had been surrounding the device. The Device-Capture Histology method, presented here, overcomes many limitations of the conventional Device-Explant Histology method, by collecting the device and surrounding tissue intact for subsequent labeling. With the implant remaining in situ, accurate and precise imaging of the morphologically preserved tissue at the brain/microdevice interface can then be collected and quantified. First, this article presents the Device-Capture Histology method for obtaining and processing the intact, undisturbed microdevice-tissue interface, and imaging using fluorescent labeling and confocal microscopy. Second, this article gives examples of how to quantify features found in the captured peridevice tissue. We also share histological data capturing (1) the impact of microdevice implantation on tissue, (2) the effects of an experimental anti-inflammatory coating, (3) a dense grouping of cell nuclei encapsulating a long-term implant, and (4) atypical oligodendrocyte organization neighboring a long term implant. Data sets collected using the Device-Capture Histology method are presented to demonstrate the significant advantages of processing the intact microdevice-tissue interface, and to underscore the utility of the method in understanding the effects of the brain-implantable microdevices on nearby tissue.
准确评估脑植入微器件的生物兼容性仍然是一个艰巨的挑战。目前的组织学方法要求在组织处理前提取器件,这通常会破坏和移除器件周围感兴趣的组织。本文提出的器件捕获组织学方法克服了传统器件外植组织学方法的许多局限性,它可以完整地收集器件和周围组织,以便后续进行标记。由于植入物仍处于原位,因此可以对脑/微器件界面处形态保存完好的组织进行精确和准确的成像和定量分析。首先,本文介绍了用于获取和处理完整、未受干扰的微器件-组织界面的器件捕获组织学方法,以及使用荧光标记和共聚焦显微镜进行成像的方法。其次,本文给出了如何量化捕获的器件周围组织中特征的示例。我们还分享了组织学数据采集(1)微器件植入对组织的影响,(2)实验性抗炎涂层的效果,(3)长期植入物周围密集的细胞核包封,以及(4)长期植入物附近异常的少突胶质细胞组织。使用器件捕获组织学方法收集的数据集用于演示处理完整的微器件-组织界面的显著优势,并强调该方法在了解脑植入微器件对附近组织影响方面的实用性。