Field Emission Scanning Electron Microscope for use in a multi-user Facility

用于多用户设施的场发射扫描电子显微镜

• 批准号: 7794699
• 负责人: LAURA SANTAMBROGIO
• 金额: $ 50万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-02-01 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7794699
• 关键词: Appearance; Arts; Bacteria; Biogenesis; Biological; Cell secretion; Cell surface; Characteristics; Cytoplasmic Granules; Dehydration; Devices; Electron Microscope; Electron Microscopy; Electrons; Freezing; Funding; Gold; Image; Investigation; Location; Lysosomes; Modification; Morphologic artifacts; Morphology; Nematoda; Organelles; Physiological; Research Personnel; Resolution; Resources; Sampling; Scanning; Scanning Electron Microscopy; Scanning Transmission Electron Microscopy Procedures; Source; Structure; Surface; System; Time; cell motility; cellular imaging; fungus; investigator training; late endosome; pathogen; receptor; voltage

DESCRIPTION (provided by applicant): High resolution morphological surface analysis is an integral part of biological investigations under both physiological and pathological conditions. The fine structure modifications induced during cell motility or cellular secretion are captured in a real time manner by scanning electron microscopy (SEM) analysis of quick-frozen cell surfaces. Similarly, surface morphological changes observed in purified sub-cellular organelles (late endosomes and lysosomes) during various experimental conditions can be powerfully visualized by SEM analysis. Surface analysis of bacteria and fungi can provide very useful information on their morphological changes under differential experimental conditions. Distinct from imaging obtained by transmission electron microscopy (TEM), the SEM analysis is capable of producing high-resolution images that have great depth of field yielding a characteristic three-dimensional appearance useful for understanding the fine surface structure of a sample. For this major reason both TEM and SEM should often both be applied in the investigation of ultrastructural morphology of biological samples. This is a resubmission application for funding of a state of the art high resolution scanning electron microscope and an associated cryotransfer system. The requested SEM will be part of the Analytical Imaging Facility (AIF), a multi user imaging resource, and available to all trained investigators. Currently, a much out of date JEOL JSM6400 SEM installed 1989, is present in the facility which does not fulfill the needs of these investigators. A new SEM is required to image high magnification (>20,000x) at low accelerating voltage (1-2 kV) with a high brightness electron source for detecting 10 nm gold with backscatter electrons. A cryotransfer station will enable imaging of hydrated samples to avoid the artifacts of dehydration. The common scientific need of the major user in this application is the ability to obtain high resolution surface imaging from cells, organelles or pathogens under different experimental conditions. The cryo field emission scanning electron microscope described in this application will allow the assembled major users to analyze fine structural aspect of endosomal biogenesis and surface receptor distribution; pathogen surface morphology, receptor location and elemental distribution; nanofabricated devices; nematode morphology; and granule secretion which will be otherwise precluded.

描述（由申请人提供）：高分辨率形态表面分析是生理和病理条件下生物学研究的一个组成部分。通过对速冻细胞表面的扫描电镜（SEM）分析，可以实时捕捉到细胞运动或细胞分泌过程中引起的细微结构变化。同样，在不同的实验条件下，纯化的亚细胞器（晚期内体和溶酶体）的表面形态变化可以通过扫描电镜分析有力地观察到。细菌和真菌的表面分析可以为它们在不同实验条件下的形态变化提供非常有用的信息。与通过透射电子显微镜（TEM）获得的成像不同，扫描电镜分析能够产生高分辨率的图像，这些图像具有很大的景深，产生特征的三维外观，有助于理解样品的精细表面结构。由于这一主要原因，透射电镜和扫描电镜通常都应应用于研究生物样品的超微结构形态。这是一个高分辨率扫描电子显微镜和相关冷冻转移系统的再提交申请。所要求的扫描电镜将是分析成像设备（AIF）的一部分，这是一个多用户成像资源，可供所有训练有素的研究人员使用。目前，1989年安装的JEOL JSM6400 SEM已经过时，无法满足这些调查人员的需要。在低加速电压（1-2 kV）下，使用高亮度电子源对10 nm金进行背散射检测，需要一种新的扫描电镜（SEM）进行高放大成像（bbb20 000倍）。冷冻转移站将使水合样品成像，以避免脱水的伪影。在此应用中，主要用户的共同科学需求是在不同实验条件下从细胞，细胞器或病原体获得高分辨率表面成像的能力。本应用中描述的冷场发射扫描电子显微镜将允许组装的主要用户分析内体生物发生和表面受体分布的精细结构方面；病原体表面形态、受体位置和元素分布；nanofabricated设备;线虫形态;和颗粒分泌，否则将被排除在外。