Staining Protocols for Human Pancreatic Islets
Staining Protocols for Human Pancreatic Islets
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DOI：
10.3791/4068
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发表时间：
2012-05-01
影响因子：
1.2
通讯作者：
Schneider, Lynda
中科院分区：
文献类型：
--
作者：
Campbell-Thompson, Martha L.;Heiple, Tiffany;Schneider, Lynda
成人胰腺中胰岛面积和数量以及内分泌细胞组成的估计值从几十万到几百万不等，β质量范围从500到1500 mg（1 - 3）。有了这种已知的异质性，一个标准的处理和染色程序被开发出来，以便胰腺区域被清楚地定义，胰岛的特点是使用严格的组织病理学和免疫定位examinations.Standard程序处理从器官供体回收的人胰腺描述在本系列的第1部分。将胰腺加工成3个主要区域（头部、身体、尾部），然后进行横切。胰头的横切面根据大小进一步划分，并按顺序编号以表示子部分。这种标准化允许一个完整的横截面分析的头部区域，包括钩状区域，其中包含主要由胰腺多肽细胞组成的胰岛尾region.The目前的报告包括本系列的第2部分，并描述了用于连续切片和胰腺石蜡切片的组织病理学特征的程序，重点是胰岛内分泌细胞，复制和T细胞浸润。胰腺切片的病理学旨在表征外分泌、导管和内分泌成分。评价外分泌室是否存在胰腺炎（活动性或慢性）、萎缩、纤维化和脂肪，以及导管系统，特别是与胰腺导管内瘤形成的关系（4）。使用H & E和IHC染色评估胰岛的形态、大小和密度、内分泌细胞、炎症、纤维化、淀粉样蛋白以及复制或凋亡细胞的存在。第2部分中描述的最后一个组成部分是提供染色的载玻片作为数字化的全载玻片图像。数字化的切片在在线病理学数据库中按病例和胰腺区域组织，创建虚拟生物库。目前，参与1型糖尿病研究的200多名临床医生和科学家可以访问该在线收集。在线数据库提供了一种快速和完整的数据共享手段，并为研究人员选择石蜡或冷冻连续切片的块。
Estimates of islet area and numbers and endocrine cell composition in the adult human pancreas vary from several hundred thousand to several million and beta mass ranges from 500 to 1500 mg(1-3). With this known heterogeneity, a standard processing and staining procedure was developed so that pancreatic regions were clearly defined and islets characterized using rigorous histopathology and immunolocalization examinations.Standardized procedures for processing human pancreas recovered from organ donors are described in part 1 of this series. The pancreas is processed into 3 main regions (head, body, tail) followed by transverse sections. Transverse sections from the pancreas head are further divided, as indicated based on size, and numbered alphabetically to denote subsections. This standardization allows for a complete cross sectional analysis of the head region including the uncinate region which contains islets composed primarily of pancreatic polypeptide cells to the tail region.The current report comprises part 2 of this series and describes the procedures used for serial sectioning and histopathological characterization of the pancreatic paraffin sections with an emphasis on islet endocrine cells, replication, and T-cell infiltrates. Pathology of pancreatic sections is intended to characterize both exocrine, ductular, and endocrine components. The exocrine compartment is evaluated for the presence of pancreatitis (active or chronic), atrophy, fibrosis, and fat, as well as the duct system, particularly in relationship to the presence of pancreatic intraductal neoplasia(4). Islets are evaluated for morphology, size, and density, endocrine cells, inflammation, fibrosis, amyloid, and the presence of replicating or apoptotic cells using H&E and IHC stains.The final component described in part 2 is the provision of the stained slides as digitized whole slide images. The digitized slides are organized by case and pancreas region in an online pathology database creating a virtual biobank. Access to this online collection is currently provided to over 200 clinicians and scientists involved in type 1 diabetes research. The online database provides a means for rapid and complete data sharing and for investigators to select blocks for paraffin or frozen serial sections.