Intrinsic Fluorimetric Imaging at the Cellular and System Levels

细胞和系统水平的本征荧光成像

• 批准号: 7230189
• 负责人: ANDREW T SORNBORGER
• 金额: $ 17.87万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7230189
• 关键词: Action Potentials; Animal Model; Animals; Behavior; Benchmarking; Biological Models; Blood; Blood capillaries; Blood flow; Brain; Calcium Spikes; Cell Respiration; Cells; Coenzymes; Complex; Cultured Cells; Data; Detection; Development; Embryo; Event; Eye; Flavoproteins; Fluorescence; Functional Imaging; Generations; Goals; Hippocampus (Brain); Human; Image; Imaging Device; Imaging technology; In Vitro; Larva; Life; Link; Measurement; Measures; Metabolic; Methods; Microscopy; Motivation; NADH; Neurons; Neurosciences; Nicotinamide adenine dinucleotide; Noise; Organism; Oxidation-Reduction; PC12 Cells; Physiological; Preparation; Problem Solving; Proxy; Relative (related person); Research; Research Personnel; Resolution; Respiration; Sensory; Signal Transduction; Standards of Weights and Measures; Statistical Data Interpretation; Stretching; Structure of trigeminal ganglion; System; Techniques; Tissues; Vertebrates; Zebrafish; base; capillary; day; design; experience; fluorophore; in vivo; neural circuit; pressure; programs; relating to nervous system; response; tool; two-photon

DESCRIPTION (provided by applicant): This research is designed to develop a new tool for measuring functional neural activity from the sub-cellular to the system scale. Using two-photon microscopy of the NADH/FP signal in vitro in cultured PC12 cells and in vivo in zebrafish, we will establish the important neuronal, activation-related metabolic events that cause modulations in the fluorimetric signal. In cultured PC12 cells, we will establish the spatio-temporal signature of known cellular physiological events such as stretch, pressure change, action potential generation and calcium spikes in benchmark, two-photon imaging studies giving important information on the relative amplitudes and durations of the metabolic events that we will use as proxies for the measurement of neuronal electrical and signaling activity. Using zebrafish, we will bring our imaging effort to the organismal level. We will measure intrinsic NADH/FP signal modulation in the well-characterized trigeminal ganglion. By comparing the NADH/FP signal with standard indicators of neuronal activity, we will characterize the spatio-temporal NADH/FP signal in a live preparation determining the detailed dynamics of the signal and determining differences with the signal arising from cultured cells. Because of the relatively low fluorescence amplitudes of NADH and FP relative to extrinsic fluorophores, the signal-to-noise ratio in two-photon NADH/FP imaging measurements limits the accuracy of quantitative measurements. Therefore, we will develop multivariate statistical analysis methods for estimating the multichannel NADH and FP signals. This will allow us to more accurately detect and quantitate metabolic changes in intact tissue. The resulting methods will be generally applicable to other dual-channel, spectroscopic signals.

描述（由申请人提供）：本研究旨在开发一种新的工具，用于测量从亚细胞到系统尺度的功能性神经活动。在体外培养的PC 12细胞和斑马鱼体内使用NADH/FP信号的双光子显微镜，我们将建立重要的神经元，激活相关的代谢事件，导致荧光信号的调制。在培养的PC 12细胞中，我们将建立已知的细胞生理事件的时空特征，例如在基准、双光子成像研究中的拉伸、压力变化、动作电位产生和钙峰，提供关于代谢事件的相对振幅和持续时间的重要信息，我们将使用这些信息作为神经元电和信号传导活动的测量的代理。利用斑马鱼，我们将把我们的成像努力带到生物体水平。我们将测量固有的NADH/FP信号调制的特征良好的三叉神经节。通过将NADH/FP信号与神经元活动的标准指标进行比较，我们将表征活体制剂中的时空NADH/FP信号，确定信号的详细动力学并确定与培养细胞产生的信号的差异。由于相对于外来荧光团而言，NADH和FP的荧光振幅相对较低，因此双光子NADH/FP成像测量中的信噪比限制了定量测量的准确性。因此，我们将发展多元统计分析方法来估计多通道NADH和FP信号。这将使我们能够更准确地检测和定量完整组织中的代谢变化。所得到的方法将普遍适用于其他双通道，光谱信号。

项目成果

Estimating weak ratiometric signals in imaging data. II. Meta-analysis with multiple, dual-channel datasets.

• DOI: 10.1364/josaa.25.002185
• 发表时间: 2008-09
• 期刊: Journal of the Optical Society of America. A, Optics, image science, and vision
• 作者: A. Sornborger;J. Broder;A. Majumder;Ganesh Srinivasamoorthy;E. Porter;Sean S Reagin;Charles Keith;J. Lauderdale

A multivariate, multitaper approach to detecting and estimating harmonic response in cortical optical imaging data.

• DOI: 10.1016/j.jneumeth.2011.09.018
• 发表时间: 2012-01-15
• 期刊: JOURNAL OF NEUROSCIENCE METHODS
• 影响因子: 3
• 作者: Sornborger, A. T.;Yokoo, T.
• 通讯作者: Yokoo, T.