LEICA TCS SP2 ADBS CONFOCAL MICROSCOPE

描述（由申请人提供）：申请要求支持徕卡TCS SP2 AOBS光谱共聚焦显微镜。最近推出的频谱可编程徕卡AOBS技术允许无滤波器操作，这增加了信号强度，从而减少了光漂白和延长了电池寿命。该显微镜将被纽约大学医学院（NYU SOM）微生物学系（4个专业和1个次要使用者）和生物化学系（3个专业和2个次要使用者）的成员共享使用。在他们之间，用户有18个NIH拨款。这些研究人员提出的研究主要表征了正常和病毒感染或应激细胞中调节细胞生长的信号通路的组成部分。这些项目包括确定活细胞和固定细胞中蛋白质的亚细胞定位，以及使用FRAP分析活细胞中的核质穿梭和使用FRET分析多种蛋白质因子的相互作用。主要使用者的研究项目包括：i)疱疹病毒裂解和潜伏基因表达的转录控制（Wilson）， ii)疱疹病毒感染细胞中Us11 mRNA翻译的调节（Mohr）， iii)在感染细胞核中维持非复制的乙型肝炎病毒DNA片段（Schneider）， iv)通过核蛋白-p53相互作用调节p53稳定性的新机制的表征（Borowiec）， v)由RP2缺陷引起的视网膜变性的机制。视网膜色素变性的一个致病因素（Cowan）， vi)糖皮质激素受体磷酸化异构体的定位（Garabedian），以及vii) Kv4亚家族的K+通道蛋白的转运及其与相关蛋白相互作用的动力学（Rudy）。目前，由于纽约大学SOM的共聚焦设备严重供不应求，使用受到严重限制，这些研究无法取得重大进展。因此，这些美国国立卫生研究院支持的项目的成功需要全天候使用高分辨率共聚焦显微镜。作为额外的好处，该仪器将加强对两个系的学生和博士后研究员的培训，并在用户群体之间提供更大的协同作用，从而产生新的研究方向。该部主席和用户团体已承诺为仪器维修和培训提供足够的资金。

DESCRIPTION (provided by applicant): The application requests support for a Leica TCS SP2 AOBS Spectral Confocal Microscope. The recently introduced spectrally programmable Leica AOBS technology allows a filter-free operation, which increases the signal strength allowing reduced photobleaching and longer cell lifetimes. The microscope will be a shared facility used by members of the Departments of Microbiology (4 major and 1 minor users) and Biochemistry (3 major and 2 minor users) at New York University School of Medicine (NYU SOM). Between them, the users have 18 NIH grants. The proposed studies of these investigators primarily characterize the components of signaling pathways that regulate cell growth in normal and virally-infected or stressed cells. These projects involve the determination of sub-cellular localization of proteins in living and fixed cells, and analysis in living cells of nucleocytoplasmic shuttling using FRAP and interactions of multiple protein factors using FRET. The projects of the major users include studies of: i) transcriptional control of lytic and latent gene expression in herpesviruses (Wilson), ii) modulation of mRNA translation by Us11 in herpesvirus-infected cells (Mohr), iii) maintenance of a non-replicating Hepatitis B virus DNA episome in the infected cell nucleus (Schneider), iv) characterization of a novel mechanism for regulating p53 stability through a nucleolin-p53 interaction (Borowiec), v) the mechanism of retinal degeneration caused by defects in RP2, a causative factor in retinitis pigmentosa (Cowan), vi) localization of glucocorticoid receptor phosphorylation isoforms (Garabedian), and vii) trafficking of K+ channel proteins of the Kv4 subfamily and the dynamics of their interactions with associated proteins (Rudy). Significant progress towards these studies is currently prevented because confocal facilities at NYU SOM are heavily over-subscribed and access is severely limited. The success of these NIH-supported projects therefore requires full-time access to a high-resolution confocal microscope. As additional benefits, the instrument will enhance the training of students and postdoctoral fellows in the two departments, and provide increased synergy between user groups that will generate new research directions. The Department chairs and user groups have committed sufficient funds for instrument maintenance and training.