High resolution fluorescence stereomicroscope for 3D imaging

用于 3D 成像的高分辨率荧光体视显微镜

• 批准号: 345750-2007
• 负责人: Matton, Daniel
• 金额: $ 3.1万
• 依托单位: Université de Montréal
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments - Category 1 (<$150,000)
• 财政年份: 2006
• 资助国家: 加拿大
• 起止时间: 2006-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=325595
• 关键词: resolution; fluorescence; stereomicroscope; 3D; imaging

In modern cell and molecular biology laboratories, careful observations of protein localization and cellular phenotypes are intensively used to gain insight into the role of specific genes in an organism. Therefore, in recent years, optical microscopy techniques have experienced a remarkable comeback in life science research. When dealing with larger specimens, upright or inverted microscopes are not adequate to provided sufficient depth of field. For this, researchers need a high resolution stereomicroscope. Furthermore, with the advent of various fluorescent protein tags, the availability of a high resolution fluorescent stereomicroscope will enable us to detect with higher efficiency protein fusions expressed in larger specimens, like pollen tubes or ovules. The equipment proposed here is intended firstly to cover our increasing and still unmet needs in: 1) dissection and observation of cleared tissues from mutants; 2) fluorescence applications that do not involve elaborate reconstructions through software analysis but that demand an increased depth of field like GFP fluorescent or aniline blue stained pollen tubes in fertilization studies; 3) higher throughput fast localization of GFP or YFP tagged protein in localization or protein-protein interaction studies. This will be particularly important for the rapid localization of cells expressing the fluorescent-tagged fusion proteins following transient expression through micro particle bombardment of onion epidermis or leaf agroinfiltration; 4) transmitted, reflected, or fluorescent light stereomicroscopy of larger samples that cannot be observed as a whole in a normal upright or inverted microscope due to poor depth of field, even at very low magnification.

在现代细胞和分子生物学实验室中，对蛋白质定位和细胞表型的仔细观察被广泛用于深入了解生物体中特定基因的作用。因此，近年来，光学显微镜技术在生命科学研究中经历了显着的回归。当处理较大的标本时，直立或倒置的显微镜不足以提供足够的景深。为此，研究人员需要高分辨率的立体显微镜。此外，随着各种荧光蛋白标签的出现，高分辨率荧光体视显微镜的可用性将使我们能够以更高的效率检测在较大标本中表达的蛋白融合体，如花粉管或胚珠。本文提出的设备首先旨在满足我们在以下方面日益增长且仍未满足的需求：1）解剖和观察来自突变体的透明组织; 2）荧光应用，其不涉及通过软件分析的精细重建，但需要增加景深，如GFP荧光或苯胺蓝染色的花粉管在受精研究中; 3）在定位或蛋白质-蛋白质相互作用研究中GFP或YFP标记的蛋白质的更高通量快速定位。这对于表达荧光标记的融合蛋白的细胞在通过洋葱表皮的微粒轰击或叶农杆菌渗透瞬时表达后的快速定位将是特别重要的; 4）较大样品的透射、反射或荧光立体显微镜，所述较大样品由于景深差而不能在正常直立或倒置显微镜中作为整体观察，即使在非常低的放大倍数下也是如此。