Optical Scatter Microscopy

光学散射显微镜

• 批准号: 6328515
• 负责人: Nada Boustany
• 金额: $ 12.26万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6328515
• 关键词: apoptosis; bioimaging /biomedical imaging; cytology; fluorescence; green fluorescent proteins; light microscopy; light scattering; mitochondria; optics; technology /technique development

DESCRIPTION (Adapted from the applicant's abstract): Although alterations in the shape and size of subcellular organelles are important markers of cell death, these important morphological changes cannot be directly quantified in living cells by conventional light microscopy. Quantitative light scattering provides an appealing solution to this problem. This technique is noninvasive, and is sensitive to the dimensions of particles with size on the order of the wavelength. Existing light scattering methods, which have been successfully utilized in flow cytometry, are spectroscopic in nature and do not permit sample imaging. Preserving spatial information is crucial to correlating the source of the scatter changes with a given cell or organelle. To this end, the investigators propose to combine light scattering spectroscopy with imaging. This novel technology, OSM, will produce the first images in which alterations in organelle morphology can be tracked in living cells in real time. The preliminary data attest to the feasibility and sensitivity of OSM. Notably, the investigators were able to detect an intracellular optical scatter change early during apoptosis, long before it was discernable by other light microscopic methods, such as DIC. However, the biological events responsible for this scatter change remain to be identified. Elucidating the relationship between the optical scatter changes and cellular activity is the principal challenge in developing OSM. If this challenge is met, OSM has the potential to transform significantly the approach to non-invasive monitoring of intracellular activity, and holds promise to fundamentally enhance high resolution light microscopy. In this proposal, the investigators present a rational strategy and step-by- step approach to implement and validate OSM. In this initial phase of development, the investigators will use OSM to study mitochondrial dysfunction during cell death. The long-term goal is to develop OSM as a tool for functional microscopy.

描述(改编自申请人的摘要)：尽管在 亚细胞器的形状和大小是细胞的重要标志 死亡，这些重要的形态变化不能直接量化 用常规光学显微镜观察活细胞。定量光散射 为这个问题提供了一个有吸引力的解决方案。这项技术是 非侵入性，并且对颗粒的尺寸很敏感 波长的顺序。现有的光散射方法，已经被 成功地用于流式细胞术，具有光谱性质和 不允许进行样品成像。保存空间信息对于 将散射变化的来源与给定的细胞或细胞器相关联。 为此，研究人员建议将光散射 光谱学与成像。这项名为OSM的新技术将生产出第一个 可以在活体中跟踪细胞器形态变化的图像 实时检测细胞。 初步数据验证了OSM方法的可行性和敏感性。 值得注意的是，研究人员能够检测到细胞内的光学 在细胞凋亡过程中早期的散布变化，早在它被其他人发现之前 光学显微镜方法，如DIC。然而，生物事件 造成这种分散变化的原因仍有待确定。澄清 光散射变化与细胞活动之间的关系是 发展OSM的主要挑战。如果这一挑战得到满足，OSM将拥有 显著改变非侵入性监测方法的潜力 细胞内活性，并有望从根本上提高高 分辨率光学显微镜。 在这份提案中，调查人员提出了一种理性的策略和循序渐进的- 实施和验证OSM的分步方法。在这一初始阶段， 开发，研究人员将使用OSM来研究线粒体功能障碍 在细胞死亡期间。我们的长期目标是将OSM开发为 功能显微镜。