Morphology and Histology Core

项目摘要-核心B 该计划项目的三个项目在很大程度上依赖于使用鼠标模型来演示其 假设。因此，形态和组织学核心的目的是巩固设备和 为这些项目的研究人员提供最先进的、质量受控的组织学和 骨、胰腺和脑标本的组织形态计量学分析以及显微CT和生物力学 对老鼠骨骼的分析。对此计划项目的核心贡献将分为5个级别： 1.对小鼠标本进行组织学和组织形态计量学研究。固定和部分脱水的小鼠 骨骼或胰腺将由项目调查人员提供给核心。然后这些样品将被 包埋在塑料介质(椎骨和长骨)或石蜡(胰腺)中。固定的小鼠脑 标本将被提供给岩芯，并被切片而不包埋或冷冻包埋 冰冻切片。 2.对小鼠组织切片进行特异性染色和免疫染色。经典组织学染色 如苏木素/曙红、甲苯胺蓝、冯·科萨/凡·吉森、Goldner三色、甲酚紫等 以及更具体的反应，如TUNEL和陷阱染色，将由核心进行 人员应调查人员的要求。核心还将进行免疫组织化学研究，使用 荧光法和酶法检测。 3.对小鼠骨、胰腺和脑标本进行组织形态计量学分析。这个 将使用OsteoMeasure组织形态测量系统(OsteoMetrics Inc.，Decatur，GA)来评估静态 骨体积、骨小梁数量和细胞计数等参数，以及骨的动力学参数 从未脱钙的骨切片中形成矿物质，如矿物质沉积率。它还将被用来 对胰腺细胞质量和脑组织切片中TUNEL或BrdU阳性细胞进行定量。将使用图像J 用于所有其他量化。核心将为调查人员提供组织形态测量数据，并将帮助 它们在评估它们的生物学相关性方面的作用。 4.对活体小鼠和小鼠骨标本进行显微CT分析。麻醉小鼠或固定的和部分的 项目调查人员将向Core提供脱水的小鼠长骨和脊椎。这些 然后将使用VivaCT 40微型CT扫描样本(瑞士Sanco Medical)。 5.对小鼠骨标本进行生物力学测试。速冻小白鼠长骨和脊椎 将由项目调查员提供给核心。然后对这些样本进行压缩 三点弯曲试验。

Project Summary – Core B The three projects of this Program Project rely heavily on the use of mouse models to demonstrate their hypotheses. The purpose of the Morphology and Histology Core is therefore to consolidate equipment and expertise to provide the Investigators of these projects with state-of-the-art, quality controlled histologic and histomorphometric analyses of bone, pancreas and brain specimen as well as microCT and biomechanical analyses of mouse bones. The Core contribution to this Program Project will be at 5 levels: 1. Processing mouse specimens for histology and histomorphometry. Fixed and partially dehydrated mouse skeletons or pancreata will be provided to the Core by project investigators. These samples will then be embedded in plastic medium (vertebrae and long bones) or paraffin (pancreas). Fixed mouse brain specimen will be provided to the Core and either be sectioned without embedding or cryo-embedded and cryo-sectioned. 2. Performing specific stainings and immunostainings of mouse tissues sections. Classic histology stainings such as Hematoxylin/Eosin, Toluidine Blue, von Kossa/Van Gieson, Goldner trichrome, Cresyl violet as well as more specific reactive ones such as TUNEL and TRAP stainings will be performed by the Core personnel at the request of investigators. The Core will also perform immunohistochemistry studies using fluorescent and enzymatic detection. 3. Performing histomorphometric analyses of mouse bone, pancreas and brain specimens. The OsteoMeasure Histomorphometry System (OsteoMetrics Inc., Decatur, GA) will be used to assess static parameters, such as bone volume, trabecular number and cell counts, and dynamic parameters of bone formation, such as mineral apposition rates, from non-decalcified bone sections. It will also be used to quantify -cell mass in pancreas and TUNEL or BrdU positive cells in brain sections. Image J will be used for all other quantifications. The Core will provide investigators with histomorphometric data and will help them in evaluating their biological relevance. 4. Analyzing live mice and mouse bone specimen by microCT. Anesthesized mice or fixed and partially dehydrated mouse long bones and vertebrae will be provided to the Core by project investigators. These specimens will then be scanned using a VivaCT 40 microCT (Scanco Medical, Switzerland). 5. Performing biomechanical tests on mouse bone specimen. Flash frozen mouse long bones and vertebrae will be provided to the Core by project investigators. These samples will then be subjected to compression and 3-point bending tests.