Leica LMD7000 Laser Microdissection System

描述（由申请人提供）：加州大学旧金山分校欧内斯特·加洛诊所和研究中心（EGCRC）的组织学和显微镜核心设施提供共享设备和培训方法，使用显微镜研究药物和酒精依赖背后的大脑分子和结构变化。这些问题传统上是通过免疫细胞化学和原位杂交来解决的，在过去的十年里，美国国立卫生研究院开发了激光捕获显微解剖（LCM）方法，该方法提供了从反染色组织学切片中选择性收集特定细胞群的可能性。原始的基于红外（IR）激光的PixCell IIe系统（Arcturus）在2001年被EGCRC购买。从那时起，该仪器使我们的研究人员能够通过RT-PCR和微阵列技术测量无脊椎动物和哺乳动物细胞亚群中的mRNA丰度。然而，在过去的几年里，这种基于红外激光的LCM设计的技术缺陷已经变得明显。首先，它只能使用5-8微米厚的切片，因此仅限于分析少量组织。其次，我们的PixCell IIe系统是手动的，不能使用自动功能勾勒出大脑区域的轮廓，然后解剖出大脑区域。此外，为了获得最有效的红外脉冲，必须在组织收集之前和收集过程中重新聚焦手动激光，这非常耗时。此外，由于需要从每只动物身上收集成千上万的细胞，研究被病毒感染的大脑区域以进行RNA干扰研究已被证明是非常困难的。因此，尽管在我们的实验中对LCM有很大的需求，但我们目前的系统是非常不足的。最近，LCM供应商开发了使用紫外线（UV）激光器作为切割工具的系统，这一发展已被证明是一项重大改进。由于我们的PixCell IIe是不可升级的，我们现在寻求获得最新的仪器，从徕卡微系统的LDM7000。所要求的仪器将配备高效紫外切割激光器，自动区域采集和其他选项，这将大大有助于以更低的成本和时间获得更好的样品。有了LDM7000，我们的研究人员将能够继续进行美国国立卫生研究院资助的实验，避免了目前EGCRC过时设备所带来的严重限制。

DESCRIPTION (provided by applicant): The Histology and Microscopy Core facility at the Ernest Gallo Clinic and Research Center (EGCRC) at the University of California, San Francisco provides shared equipment and training in methods that use microscopy for investigating molecular and structural changes in the brain that underlie drug and alcohol dependence. These questions have been traditionally addressed using immunocytochemistry and in situ hybridization, and for the past decade, with the NIH-developed method of laser capture microdissection (LCM), which offers the possibility of selectively collecting specific cell populations from counterstained histological sections. The original infrared (IR) laser-based PixCell IIe system (Arcturus) was purchased by the EGCRC in 2001. Since then this instrument has allowed our investigators to measure mRNA abundance in subpopulations of invertebrate and mammalian cells by RT-PCR and microarray technology. However, over the last several years, technical drawbacks with this IR laser-based LCM design have become evident. First, it can optimally use only 5-8 micron thick sections and is thus limited to analysis of small amounts of tissue. Second, our PixCell IIe system is manual and cannot outline and then dissect out a brain region using an automated function. Furthermore, to obtain the most effective IR pulses, one must refocus the manual laser before and during tissue collection, which is very time-consuming. In addition, investigating regions of the brain infected with viruses for RNA interference studies has proven very difficult to perform due to the need to collect many thousands of cells from each animal. Therefore, despite a significant need for LCM in our experiments, our current system is extremely inadequate. Recently, LCM vendors have developed systems that use ultraviolet (UV) lasers as a cutting tool and this development has proven to be a major improvement. Since our PixCell IIe is not upgradeable, we now seek to acquire the most up-to-date instrument, an LDM7000 from Leica Microsystems. The requested instrument will be equipped with the highly effective UV cutting laser, automated region collection and other options that will greatly assist in obtaining better samples at reduced cost and time. With the LDM7000 our investigators will be able to continue to perform NIH-funded experiments and avoid the serious limitations imposed by the outdated equipment currently at the EGCRC.