Confocal High Speed Imaging System

描述（由申请人提供）：本申请申请资金用于购买用于共聚焦荧光成像，亚细胞光解和单细胞膜片钳控制的高速成像系统。该共聚焦成像系统将用于马里兰大学生物技术研究所（UMBI）的一个新的共享仪器设施。用户已经对系统的每个元素进行了广泛的测试，并在拟议的项目中概述了使用这种新型的最先进的成像系统的计划工作。成像系统包括三个部分。共聚焦仪器是蔡司生命仪器，能够以1 kHz 50行512像素或全帧（512 X 512）以120 Hz（每张图像8.3 ms）的帧速率获取XY图像。可以在62.5 kHz（16微秒）下获得直线扫描图像（1 X 512）。该系统的光解组件以Spectra Physics的hipo -355 OEM激光器为中心，运行频率为100 kHz，平均功率为5 W。扫描头可以放置在倒置显微镜上进行最佳单细胞膜片钳实验，也可以放置在直立显微镜上进行脑切片或特殊培养条件。触发和激光发射光学是定制的，用于蔡司Live5仪器和膜片钳电子设备。它可以同步到微秒级精度，这足以满足所有计划的工作。该系统的膜片钳组件以HEKA EPC10膜片钳单元和运行Patchmaster采集软件的LIH-1600计算机接口为中心。巴尔的摩马里兰大学系统（UMBI和UMB）的研究人员在活细胞中高时间和空间分辨率的钙信号研究方面取得了30年的声誉。他们在心脏、骨骼和平滑肌中发现了Ca2+火花，并研究了这些组织在正常和病理条件下的功能，在该领域处于世界领先地位。他们在神经元和脑切片成像方面也有类似的声誉。这种提议的新仪器在巴尔的摩-华盛顿地区的任何地方都没有，它将使这些科学家能够进行资助的研究并取得重大进展。

DESCRIPTION (provided by applicant): This application requests funds to purchase a high speed imaging system for confocal fluorescence imaging, subcellular photolysis and patch clamp control of single cells. The confocal imaging system will be used in a new shared instrument facility at the University of Maryland Biotechnology Institute (UMBI). Extensive testing of each element of the system by the users has been carried out and planned work using this novel state-of-the-art imaging system is outlined in the proposed projects. The imaging system involves three components. The confocal instrument is the Zeiss LiveS instrument that is capable of acquiring XY images at 1 kHz at 50 lines by 512 pixels or full frame (512 X 512) at a frame rate of 120 Hz (8.3 ms per image). Linescan images (1 X 512) can be obtained at 62.5 kHz (16 microseconds). The photolysis component of the system is centered on a Spectra Physics HIPPO-355 OEM laser running at 100 kHz with an average power of 5 W. The scan head can be placed on an inverted microscope for optimal single cell patch clamp experiments or on an upright microscope of brain slice or special culturing conditions. The triggering and laser launch optics are customized for use with both the Zeiss Live5 instrument and with the patch clamp electronics. It can be synchronized to microsecond precision, which is adequate for all planned work. The patch clamp component of the system is centered on a HEKA EPC10 patch clamp unit and LIH-1600 computer interface running Patchmaster acquisition software. Researchers at the University System of Maryland in Baltimore (UMBI and UMB) have achieved a three-decade reputation for the study of calcium signaling at high temporal and spatial resolution in living cells. They have discovered Ca2+ sparks in cardiac, skeletal and smooth muscle, have investigated function in these tissues in normal and pathological conditions and have been world- wide leaders in this area. They have a similar reputation in neuronal and brain slice imaging. This proposed new instrumentation is not available anywhere in the Baltimore-Washington area and will allow these scientists to carry out funded research and to make significant advances.