Request for Ultracryomicrotomy equipment

描述（由申请人提供）：本申请旨在获得资金，用于购买两台带冷冻装置的超微切片机，用于匹兹堡大学生物成像中心（CBI）的EM设施。免疫电子显微镜，使用冷冻切片进行免疫金标记，是CBI提供的一项基本服务，该中心自1991年首次开始运营以来，一直在连续使用两台超微冷冻切片机。最初，cryokits都是基于两个UltraCut E单元，一个是新的，另一个是20世纪80年代初的。当其中一个单元出现故障时，它被保留为备件，并在大约13年前被Ultracut S Cryokit替换。两年前，S单元冷冻箱发生了不可修复的故障（徕卡根本不为这些系统提供任何支持，由于冷冻箱的电子和温度控制多次故障，独立承包商无法修复该设备）。我们已经设法使用从以前退役的系统中收集的备件来保持旧的UltraCut E单元运行，但是这个系统现在已经超过16年了，并且无法可靠地修复，当然即使全天使用也无法跟上我们用户群的需求。基本上，这个系统也必须退休。我们将继续使用它们作为切片机（事实上，Ultracut S单元已经完成了这一角色），但我们必须购买2个新的冷冻系统。不幸的是，由于生产过时，这只能通过购买完整的超薄切片机和冷冻机来完成。重要的是要认识到我们确实需要两个系统，而不是简单地要求两个设备超过SIG赠款机制的资金下限。在第一个冷冻切片机失败后，我们试图为单个冷冻切片机添加一个“填充”应用程序，但我们觉得这有点过分。现在，我们处于一个没有赢家的局面，我们必须更换两个系统（校园内没有其他的超微冷冻切片机或核心研究EM设施，匹兹堡也没有正常运行的超微冷冻切片机）。这一需求在对中心使用情况的描述、个人用户的预计使用情况以及高级机构官员的支持信中得到了定义。公共卫生相关性：免疫电子显微镜使用冷冻切片进行免疫金标记，是准确确定蛋白质在细胞和组织中位置的重要工具。分辨率至少比使用光学显微镜的分辨率好10倍。这种分辨率优势使这种方法成为一种非常重要的工具，通过定义进口分子在细胞内的位置和工作来了解疾病的分子机制。

DESCRIPTION (provided by applicant): This request is for funds to purchase two Ultramicrotomes with cryokits for use in the EM facility within the Center for Biologic Imaging (CBI) at the University of Pittsburgh. Immuno-electron microscopy, using ultrathin cryosections for immunogold labeling, is an essential service provided by the CBI, and the center has had two ultracryomicrotomes in continuous use since it first started operations in 1991. Initially the cryokits were both based around two UltraCut E units, one was new the other dated from the early 1980's. When one of these units failed it was kept for spare parts and replaced with an Ultracut S Cryokit some 13 years ago. 2 years ago the S-unit cryokit failed irreparably (Leica does not provide any support for these systems at all, and independent contractors were unable to repair the device due to multiple failures of the electronics and temperature control of the cryochamber). We have managed to keep the older UltraCut E unit running using spare parts harvested from the previously retired system, however this system which is now over 16 years old is failing and also cannot be repaired reliably, and certainly even with all day usage does not keep up with the needs of our user base. Essentially this system must be retired also. We will continue to use them as microtomes (in fact the Ultracut S unit is already fullfilling this role) but it is imperative we purchase 2 new cryosystems. Unfortunately this can only be done by purchasing complete ultramicrotomes plus cryokits due to manufacturing obsolescence. It is important to realize we really do need two systems, and are not simply requesting two devices to get above the funding floor of the SIG grant mechanism. Following failure of the first cryokit we were tempted to put in a "padded" application for a single cryoultramicrotome, however we felt that would be a little egregious. Now we are in a no-win situation where we must replace two systems (there are no other ultracryomicrotomes or core research EM facilities on campus or functioning ultracryomicrotomes in all of Pittsburgh). This need is defined in the description of the usage of the Center, the projected usage by individual users and in letters of support from senior institutional officials Public Health Relevance: Immuno-electron microscopy, using ultrathin cryosections for immunogold labeling, is an essential tool in defining exactly where proteins are within cells and tissues. The resolution is at least 10 times better than that possible using light microscopy. This resolution advantage makes this method an extremely important tool in understanding the molecular mechanisms underlying disease by defining where molecules of import are made and work within cells.