Summary

2012年8月:本月在朱庇特

Published: August 01, 2012
doi:

Summary

传统的显微镜需要放大标本的镜头目标,并可能涉及众多的光学元件,如额外的目标,过滤器和镜子,折射和直接光光学传感器。朱庇特(可视化实验杂志)2012年8月发行,标志着从,奥兹坎实验室(美国加州大学洛杉矶分校),他们的镜头镜头无“片上”显微镜平台,他们已率先公布第三。

Abstract

传统的显微镜需要放大标本的镜头目标,并可能涉及众多的光学元件,如额外的目标,过滤器和镜子,折射和直接光光学传感器。 庇特(可视化实验杂志)2012年8月发行,标志着从,奥兹坎实验室(美国加州大学洛杉矶分校),他们的镜头镜头无“片上”显微镜平台,他们已率先公布第三。

奥兹坎实验室早在2009年,发表了他们的第一篇文章,这表明使用无透镜成像平台,全血内的图像细胞。这种成像技术,包括直接将样品标准摄像机的CCD或CMOS芯片一样,光电传感器阵列。然后,LED从样品上方的照明蒙上阴影的芯片,它可以承受图像处理的数字全息图记录。不仅这个无透镜成像系统允许一个非常大的领域观点(视野0.6-8厘米2),缺乏笨重的光学元件,使得微流体应用的优秀。奥兹坎组及其后续2011年的朱庇特的文章,其中荧光标记的白血细胞内的微芯片成像演示。

这个月在朱庇特,奥兹坎组镜头的显微镜平台,将它应用到线虫C.层析成像演示的进一步发展线虫 。通过改变光照的角度,使用电动旋转舞台,奥兹坎组多个全息片显示了如何生成可为样本,处理后,可结合成tomograms的。的的奥兹坎组确认援引咽管,5微米厚的结构,只是在一个片,并在三维重建截面可见,他们的蠕虫病毒的三维重建是成功的。

– 奥兹坎组的设计和组装 – 镜头成像系统结构紧凑,价格低廉的性质使它们适合点护理诊断应用程序,或者甚至基于手机显微镜可以在现场进行。

朱庇特的Ernest Gallo临床和研究中心在美国加州大学旧金山学会如何在单细胞的形态学变化,采用三维荧光显微镜和计算机分析可视化。在此过程中,细胞受体激动剂,拮抗剂加激动剂,或根本没有治疗,准备和荧光显微镜。三维图像采集与双光子激光系统的帮助下,Imaris和Matlab的计算机处理的荧光数据。

为了证明这项技术的力量,我们的作家量化HEK293细胞的单细胞表型的改变。三,尺寸之NAL形态分析表明激动剂治疗的细胞中的戏剧性变化。这些变化并不总是很明显,当使用传统的二维分析。这种技术允许在单个细胞的形态变化进行量化和特点,以药物刺激的反应,药物研发阿森纳提供了强大的新的评估工具。

本月初,在朱庇特应用物理系,我们前往南卡罗来纳州克莱姆森大学材料科学与工程学院访问。在这里,研究人员展示我们如何构建实验室研究锂离子电池。

一个锂钮扣电池的基本组成部分,是工作电极,锂电池电解液和锂箔电极。纽扣电池中的不同组件都经过精心组装和密封与压接机。然后可以完成的纽扣电池在电池测试仪的特点。使用其自制的钮扣电池,我们的研究人员可以尝试使用不同的材料,使锂钮扣电池的阴极。

这在朱庇特的免疫和感染的一个月,我们前往瑞士巴塞尔,见证寄生虫感染的诊断技术。我们也看到,调查如何执行儿童的折返跑测试,以评估这些感染对体能的影响。

寄生虫是寄生蠕虫,可以住进去人类和动物。许多物种的肠道感染,可通过土壤传播。作者展示了改良加藤技术,这是用来诊断几种常见的土壤传播的蠕虫。他们还表现出的的贝尔曼技术,它采用光刺激趋光蠕虫样品准备的某些类型的迁移。蠕虫物种的各类随后可以根据形态。

我们的作者然后说明如何建立并执行一个20米的儿童穿梭运行测试,测量蠕虫感染对体能的影响。这种方法可以帮助确定整体健康的土壤传播的蠕虫感染的真实负担。

这个简要的亮点,但将在八月公布的几个显着的视频,文章。我们还将专题文章,演示如何表达基因在鸡胚脑,研究中性粒细胞轮回衬有内皮细胞,在平行流室,并精确地测量外基质蛋白降解。

Protocol

Determining Soil-transmitted Helminth Infection Status and Physical Fitness of School-aged Children Peiling Yap1, 2, Thomas Fürst1, 2, Ivan Müller1, 2, Susi Kriemler1, 2, Jürg Utzinger1, 2, Peter Steinmann1, 21Department of Epidemiology and Public Health, Swiss Tropical and Public Health Institute, Basel, Switzerland, 2University of Basel, Basel, Switzerland Chronic infection with soil-transmitted helminths (STHs) causes malabsorption, stunting, and wasting in the growing child. Hence, it is plausible that these infections also reduce the physical fitness of children. Here, we visualize two techniques for the diagnosis of STHs and the 20-meter shuttle run test for assessing children’s physical fitness. In ovo Electroporation in Chick Midbrain for Studying Gene Function in Dopaminergic Neuron Development Ben Yang1,*, Lauren B. Geary1,*, Yong-Chao Ma1, 21Northwestern University Feinberg School of Medicine, Children’s Hospital of Chicago Research Center, 2Departments of Pediatrics, Neurology and Physiology, Northwestern University Feinberg School of Medicine* These authors contributed equally To assess the function and the regulation of genes during the development of midbrain dopaminergic neurons, we describe a method that involves in ovo electroporation of plasmid DNA constructs into embryonic chick ventral midbrain dopaminergic neuron progenitors. This technique can be used to achieve efficient expression of genes of interest to study different aspects of midbrain development and dopaminergic neuron differentiation. Isolation of Human Umbilical Vein Endothelial Cells and Their Use in the Study of Neutrophil Transmigration Under Flow Conditions Anutosh Ganguly, Hong Zhang, Ritu Sharma, Sean Parsons, Kamala D. PatelDepartment of Physiology and Pharmacology, University of Calgary This article first describes a procedure for isolating human endothelial cells from umbilical veins and then shows how to use these cells to examine neutrophil transmigration under flow conditions. By using a low-volume flow chamber made from a polymer with the optical characteristics of glass, live-cell fluorescent imaging of rare cell populations is also possible. Construction and Testing of Coin Cells of Lithium Ion Batteries Archana Kayyar1, Jiajia Huang1, Mojtaba Samiee1, Jian Luo1, 21School of Materials Science and Engineering, Clemson University, 2Center for Optical Materials Science and Engineering Technologies, Clemson University A protocol to construct and test coin cells of lithium ion batteries is described. The specific procedures of making a working electrode, preparing a counter electrode, assembling a cell inside a glovebox and testing the cell are presented. Quantitative Measurement of Invadopodia-mediated Extracellular Matrix Proteolysis in Single and Multicellular Contexts Karen H. Martin, Karen E. Hayes, Elyse L. Walk, Amanda Gatesman Ammer, Steven M. Markwell, Scott A. WeedDepartment of Neurobiology and Anatomy, Program in Cancer Cell Biology, Mary Babb Randolph Cancer Center, West Virginia University We describe the prototypical method for producing microscope coverslips coated with fluorescent gelatin for visualizing invadopodia-mediated matrix degradation. Computational techniques using available software are presented for quantifying the resultant levels of matrix proteolysis by single cells within a mixed population and for multicellular groups encompassing entire microscopic fields. Lensfree On-chip Tomographic Microscopy employing Multi-Angle Illumination and Pixel Super-Resolution Serhan O. Isikman1, Waheb Bishara1, Aydogan Ozcan1, 2, 31Electrical Engineering Department, University of California, Los Angeles , 2Bioengineering Department, University of California, Los Angeles , 3California NanoSystems Institute, University of California, Los Angeles Lensfree optical tomography is a three-dimensional microscopy technique that offers a spatial resolution of <1 μm × <1 μm × <3 μm in x, y and z dimensions, respectively, over a large imaging-volume of 15-100 mm3, which can be particularly useful for integration with lab-on-a-chip platforms. An Analytical Tool that Quantifies Cellular Morphology Changes from Three-dimensional Fluorescence Images Carolina L. Haass-Koffler1, 2, Mohammad Naeemuddin1, Selena E. Bartlett1, 31Medications Development, Ernest Gallo Clinic and Research Center, University of California, San Francisco , 2Clinical Pharmacology and Experimental Therapeutics, University of California, San Francisco , 3Translational Research Institute and the Institute for Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia We developed a software platform that utilizes Imaris Neuroscience, ImarisXT and MATLAB to measure the changes in morphology of an undefined shape taken from three-dimensional confocal fluorescence of single cells. This novel approach can be used to quantify changes in cell shape following receptor activation and therefore represents a possible additional tool for drug discovery. Lensless On-chip Imaging of Cells Provides a New Tool for High-throughput Cell-Biology and Medical Diagnostics Onur Mudanyali1, Anthony Erlinger1, Sungkyu Seo1, Ting-Wei Su1, Derek Tseng1, Aydogan Ozcan1, 21Electrical Engineering Department, University of California, Los Angeles, 2California NanoSystems Institute, University of California, Los Angeles Lensfree on-chip imaging and characterization of cells is illustrated. This on-chip cell imaging approach provides a compact and cost-effective tool for medical diagnostics and high-throughput cell biology applications, making it especially suitable for resource poor settings. Lensless Fluorescent Microscopy on a Chip Ahmet F. Coskun, Ting-Wei Su, Ikbal Sencan, Aydogan OzcanDepartment of Electrical Engineering, University of California, Los Angeles A lensless on-chip fluorescent microscopy platform is demonstrated that can image fluorescent objects over an ultra-wide field-of-view of e.g., >0.6-8 cm2 with <4μm resolution using a compressive sampling based decoding algorithm. Such a compact and wide-field fluorescent on-chip imaging modality could be valuable for high-throughput cytometry, rare-cell research and microarray-analysis.

Divulgaciones

The authors have nothing to disclose.

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Chao, W., Kolski-Andreaco, A. August 2012: This Month in JoVE. J. Vis. Exp. (66), e5016, doi:10.3791/5016 (2012).

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