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首页> 外文期刊>Bulletin of the American Physical Society >APS -2017 Annual Fall Meeting of the APS Ohio-Region Section - Event - Optimization of a nanosecond-gated spectrofluorimetry system used for the monitoring of cellular metabolism
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APS -2017 Annual Fall Meeting of the APS Ohio-Region Section - Event - Optimization of a nanosecond-gated spectrofluorimetry system used for the monitoring of cellular metabolism

机译：APS -2017年APS俄亥俄州地区分会年度秋季会议-活动-用于监控细胞代谢的纳秒级门控荧光分光光度计系统的优化

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Our lab has developed a system for the real-time monitoring of nanosecond-gated, UV-excited cellular autofluorescence (endogenous fluorescence). Time-gated detection using an intensified CCD coupled to a spectrograph enables spectral measurements at controllable time delays with respect to an excitation pulse, useful because autofluorescence is known to have many excited-state lifetime components. Under UV excitation (nitrogen laser, 337 nm wavelength, sub-ns pulse width), autofluorescence is primarily due to NAD(P)H with its ``free'' and ``protein-bound'' forms having short (extasciitilde 300 ps) and long (1--5 ns) lifetimes respectively. Time-gated detection captures the long-lifetime emission separately from the emission as a whole. Further, quantifying spectrum shape using phasor analysis allows for the assessment of two-state behavior during chemically-induced metabolic transitions, providing information at the biochemical-pathway level. Here we present an optimization of the gate timing, maximizing rejection of short-lifetime emission while maintaining contrast in spectrum-shape change during the monitoring of metabolic transitions in a model cellular system.
机译：我们的实验室开发了一种实时监控纳秒级门控，紫外线激发的细胞自发荧光（内源性荧光）的系统。使用耦合到光谱仪的增强CCD进行时间选通检测，可以在相对于激发脉冲的可控时间延迟下进行光谱测量，这是有用的，因为已知自发荧光具有许多激发态寿命分量。在紫外线激发下（氮气激光，337 nm波长，亚ns脉冲宽度），自发荧光主要归因于NAD（P）H的``游离''和``结合蛋白''形式，具有较短的时间（例如300 ps） ）和长（1--5 ns）寿命。时间门控检测从整个发射中捕获长寿命发射。此外，使用相量分析量化频谱形状可以评估化学诱导的代谢转变过程中的两态行为，从而提供生化途径水平的信息。在这里，我们介绍了门控时序的优化，在对模型细胞系统中的代谢转变进行监测期间，在维持光谱形状变化的对比度的同时，最大程度地减少了短时发射的抑制。

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