Improved Confocal Microscope: Detection & Functionality

改进的共焦显微镜：检测

• 批准号: 6932237
• 负责人: JULIAN BOREJDO
• 金额: $ 10万
• 依托单位: OPTICAL FINESSE, LLC
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-15 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6932237
• 关键词: bioengineering /biomedical engineering; bioimaging /biomedical imaging; biomedical equipment development; chemical kinetics; confocal scanning microscopy; digital imaging; fluorescence spectrometry; nanotechnology; photonics; quantum chemistry

DESCRIPTION (provided by applicant): A low-cost modification package will be developed to enhance the sensitivity of a standard confocal microscope and to upgrade its functionality as a tool for studying biological specimens. Replacing analog detectors with photon counting devices will improve signal-to-noise ratio. Digital detection through a confocal port will enable fluorescence correlation spectroscopy (PCS) to study single-molecule kinetics, diffusion, aggregation, viscosity and dynamics of cellular structures. Preliminary data obtained at moderate light levels show > 100% sensitivity enhancement with a digital detection. A much greater enhancement is expected at very low light levels. The pulse-counting electronics, new data acquisition hardware and firmware will be integrated into a compact microcontroller module with a USB connection to a PC with a Lab VIEW user interface and image analysis software. Separate circuitry to control the galvo mirrors will be devised to improve the control over the scanning process and to immobilize the galvo for the fixed-point measurements. Fixed-point measurement mode is necessary for the FCS, single-molecule spectroscopy and pharmaceutical multi-well screening. A custom-designed sideport with collimation optics will be included in the modification package to enhance the microscope collection efficiency. A low-cost fiberoptic spectrometer will be attached to the sideport to extract single molecule dynamics information from fluorescence signal (fixed-point mode) and to obtain hyperspectral image (scanning mode) of the sample. The proposed improvements are relatively simple, inexpensive and not tied to a specific brand of instrument. To facilitate a future commercialization, a microscope maintenance/modification company that is interested in distribution of the product version of the modification package has been identified.

描述（由申请人提供）：将开发一种低成本的改造包，以增强标准共焦显微镜的灵敏度并升级其作为研究生物样本的工具的功能。用光子计数装置取代模拟探测器将提高信噪比。通过共焦端口的数字检测将使荧光相关光谱（PCS）能够研究细胞结构的单分子动力学、扩散、聚集、粘度和动力学。在中等光照水平下获得的初步数据显示，数字检测的灵敏度提高了 > 100%。在非常低的光照水平下预计会有更大的增强。脉冲计数电子器件、新的数据采集硬件和固件将集成到一个紧凑的微控制器模块中，该模块通过 USB 连接到带有 LabVIEW 用户界面和图像分析软件的 PC。将设计用于控制检流镜的单独电路，以改进对扫描过程的控制并固定检流器以进行定点测量。定点测量模式是FCS、单分子光谱和药物多孔筛选所必需的。改装包中将包含带有准直光学器件的定制设计侧端口，以提高显微镜收集效率。低成本光纤光谱仪将连接到侧端口，以从荧光信号中提取单分子动力学信息（定点模式）并获得样品的高光谱图像（扫描模式）。所提出的改进相对简单、便宜并且不依赖于特定品牌的仪器。为了促进未来的商业化，已经确定了一家对分发修改包的产品版本感兴趣的显微镜维护/修改公司。