LASER SCANNING CONFOCAL MICROSCOPY FACILITY

在过去的四年里，激光扫描共聚焦显微镜设施 已在细胞血液学实验室建立。 共焦 显微镜是一种最先进的技术，可以增强对比度， 通过拒绝失焦信息来提高显微图像的分辨率。 获得样品的薄光学切片，每个图像存储在 计算机，并且图像系列被重建为立体或3- 立体图像 该设施目前用于以下用途 项目： 1. 中性粒细胞骨架和整合素分子在粘附中的作用 实力 中性粒细胞对玻璃的粘附强度在一个实验中定量。 具有渐缩间隙宽度的平行板流动室， 剪切应力梯度对强度的影响是在存在和 不存在：a）整联蛋白分子的抗体，和B）细胞松弛素D， 抑制细胞骨架蛋白F-肌动蛋白延长的试剂。 F的 肌动蛋白被荧光鬼笔环肽特异性染色， 细胞F-肌动蛋白网络用共聚焦显微镜成像。 精确 细胞厚度和F-肌动蛋白分布的测量是在 恒定剪切应力梯度的条件。 2. 白细胞迁移过程中髓样膜抗原的定位。 抗中性粒细胞膜粘附荧光单克隆抗体 分子用于检测中性粒细胞上分子的位置 在体外通过聚碳酸酯膜迁移 化学引诱物 这项研究的结果发表在1992年武装 部队病理学研究所定量组织学会议。

During the last four years, a laser scanning confocal microscopy facility has been established in the Laboratory of Cellular Hematology. Confocal microscopy is a state-of-the-art technology that enhances the contrast and resolution of microscopic images by rejecting out-of-focus information. Thin optical sections of a specimen are obtained, each image is stored on a computer, and the image series is reconstructed as a stereo or 3- dimensional image. The facility is currently used for the following projects: 1. The role of neutrophil cytoskeleton and integrin molecules on adherence strength. Adhesion strength of neutrophils to glass is quantified in a parallel plate flow chamber with a tapered gap width in which a constant shear stress gradient on the strength is evaluated in the presence and absence of: a) antibodies to integrin molecules, and b) cytochalasin D, an agent which inhibits elongation of the cytoskeletal protein, F-actin. F- actin is specifically stained with fluorescent phalloidin and the adherent cell F-actin network is imaged with the confocal microscope. Precise measurements of cell thickness and F-actin distribution are made under conditions of a constant shear stress gradient. 2. Myeloid membrane antigen location during leukocyte migration. Fluorescent monoclonal antibodies to neutrophil plasma membrane adhesion molecules are used to detect the location of the molecules on neutrophils that have migrated in vitro through polycarbonate membranes t chemoattractants. Results of this study were presented at the 1992 Armed Forces Institute of Pathology Quantitative Histopathology Conference.