DELTAVISION MULTI-MODE DECONVOLUTION MICROSCOPE

麻省理工学院生物系68号楼的6位主要研究人员提议使用高性能数字显微镜分析活细胞和固定细胞中的蛋白质分布、轴突引导、细胞运动、染色体分离和细菌细胞分裂。这项工作将使用精密三角视觉多模型显微镜（MMM）进行，该显微镜将激光扫描和基于反褶积的宽视场成像结合成一个特别强大的仪器。显微镜将被集成到一个强大的基于服务器的环境中，以便各种用户可以有效地收集和分析数据。康斯坦丁·帕顿实验室将使用麻省理工学院的MMM来代替耶鲁大学以前使用的共聚焦，以检查啮齿动物和两栖动物的活体视觉皮层中的轴突靶向。这项工作的目标是了解神经元活动如何影响轴突发育。Garrity实验室将使用显微镜检查在特定神经元中携带GFP标记的活果蝇组织中的轴突引导。分析这些神经元在不同遗传背景下的靶向作用，将揭示果蝇眼轴突引导的分子基础。格特勒实验室使用肝细胞成像技术来检测野生型和突变型小鼠细胞中基于肌动蛋白的运动性。其直接目标是确定Mena蛋白如何调节肌动蛋白细胞骨架。格罗斯曼实验室将使用MMM设施来检查枯草芽孢杆菌中参与染色体复制的蛋白质之间的空间关系。三是枯草杆菌的显著定位程度，复制蛋白和活细胞的高分辨率成像有望揭示这种定位如何随着细胞分裂周期而变化。Sinskey实验室将利用从反卷积图像中获得的空间分辨率和定量信息来研究真核真核真菌中关键PHA聚合物的生物合成。PHA对细胞的代谢具有重要意义，在可降解塑料中具有潜在的应用前景。辛斯基实验室的工作是代谢工程中先进成像技术的早期应用。Sorger实验室将利用MMM的高采集速度来捕获野生型和基因敲除小鼠和酵母细胞中染色体分离的三维延时数据。目标是揭示负责染色体分离高准确性的机制。最后，用户组将与Applied Precision公司合作，每年在麻省理工学院举办两次高级培训课程。这些对于多模型显微镜设施的长期运作至关重要，并且应该对波士顿地区的显微镜学家有普遍的好处。

Six principal investigators in Building 68 of the MIT Department of Biology, proposed to use high-performance digital microscopy to analyze protein distribution, axon guidance, cell motility, chromosome segregation and bacterial cell division in live and fixed cells. This work will be undertaken with an applied Precision DeltaVision Multi-Model Microscope (MMM) that combines laser-scanning and deconvolution- based wide field imaging into a particularly powerful instrument. The microscope will be integrated into a powerful server-based environment so that data can be effectively gathered and analyzed by various users. The Constatine Paton lab will use the MIT MMM in place of a confocal previously available at Yale to examine axon targeting in live visual cortexes fro rodents and amphibians. The goal of the work is to understand how neuronal activity affects axonal development. The Garrity lab will use the microscope to examine axon guidance in live Drosophila tissues that carry GFP markers in specific neurons. Analysis of targeting by these neurons in different genetic backgrounds will reveal the molecular basis of axon guidance in the fly eye. The Gertler lab uses liver-cell imaging to examine actin-based motility in cells derived from wild type and mutant mice. Its immediate goal is to determine how the Mena protein regulates the actin cytoskeleton. The Grossman lab will use the MMM facility is to examine the spatial relationship among proteins involved in chromosomal replication in B. subtilis. Three is a remarkable degree of localization of subtilis, replication proteins and high-resolution imaging of live cells is expected to reveal how this localization varies with the cell division cycle. The Sinskey laboratory will exploit spatially resolved and quantitative information t derived from deconvolved images to examine the biosynthesis of the critical PHA polymer in R. eutropha. PHA is of central importance to the metabolism of cells and has potential application in degradable plastics. The Sinskey lab's work is an early used of advance imaging in metabolic engineering. The Sorger Lab will use the high acquisition speed of the MMM to capture three-dimensional time lapse data on chromosome segregation in wild type and knockout mouse and yeast cells. The goal is to uncover mechanisms responsible for the high accuracy of chromosome segregation. Finally, the users group will, in conjunction with Applied Precision, present two advanced training courses per year at MIT. These will be critical for the long-term operation of the Multi-Model Microscope facility and should be of general benefit to microscopists in the Boston area.