Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA.
Department of Chemical and Systems Biology, Stanford University, Stanford, CA, 94305, USA.
Sci Rep. 2017 Jul 19;7(1):5899. doi: 10.1038/s41598-017-05614-4.
Three-dimensional tissue-structural relationships are not well captured by typical thin-section histology, posing challenges for the study of tissue physiology and pathology. Moreover, while recent progress has been made with intact methods for clearing, labeling, and imaging whole organs such as the mature brain, these approaches are generally unsuitable for soft, irregular, and heterogeneous tissues that account for the vast majority of clinical samples and biopsies. Here we develop a biphasic hydrogel methodology, which along with automated analysis, provides for high-throughput quantitative volumetric interrogation of spatially-irregular and friable tissue structures. We validate and apply this approach in the examination of a variety of developing and diseased tissues, with specific focus on the dynamics of normal and pathological pancreatic innervation and development, including in clinical samples. Quantitative advantages of the intact-tissue approach were demonstrated compared to conventional thin-section histology, pointing to broad applications in both research and clinical settings.
典型的切片组织学并不能很好地捕捉三维组织结构关系，这给组织生理学和病理学的研究带来了挑战。此外，尽管最近在成熟大脑等整个器官的完整清除、标记和成像方面取得了进展，但这些方法通常不适合用于软性、不规则和异质组织，而这些组织占绝大多数临床样本和活检组织。在这里，我们开发了一种双相水凝胶方法，该方法与自动化分析相结合，可实现对空间不规则和易碎组织结构的高通量定量体积检测。我们验证并将这种方法应用于各种发育和患病组织的检查，特别关注正常和病理胰腺神经支配和发育的动态，包括临床样本。与传统的切片组织学相比，完整组织方法具有定量优势，这表明它在研究和临床环境中有广泛的应用。
