Laser depth measurement based on time-correlated single-photon counting

A method for acquiring range data based on time-correlated single-photon counting is described. This method uses a short-pulse ( approximately 10-ps) laser diode, a detector based on a silicon single-photon avalanche diode, and standard photon-counting timing electronics. The accuracy of the technique has been measured as approximately +/-30 microm in a laboratory experiment and corresponds closely to the results of a theoretical simulation.     

新型门控光子计数法测量高散射介质的光学参量

讨论了一种新型而且简单的门控光子计数法，用来测量高散射介质的光学参量。当一束光脉冲入射到高散射介质后，该方法可以恢复出光脉冲经过散射后从介质表面逸出的波形。与传统的门控光子计数法相比，该方法利用计数器控制门可以移动的特点，通过滑动宽度为60ns的控制门实现较高的时间分辨力200ps。在此波形的基础上，Patterson—Chance方法可以快速而且准确计算出被测介质的光学参量。在模型介质上的实验，验证了该方法的可行性，实验结果与复杂的时间相关的单光子计数加曲线拟合方法相比较，误差小于10％。     

An extended scheme for counting fluorescent molecules by photon-antibunching

Acquisition of quantitative information from microscopic biological samples is highly desirable in the context of the emerging field of systems biology. We derive a statistical approach to estimate the number of fluorescent molecules in the observation volume based on a confocal microscope for single-molecule detection. The method employs ps-pulsed laser sources for excitation and time-correlated single-photon counting with 4 avalanche photon diodes (APDs) for detection of individual photons. The feasibility for estimating the number of molecules is shown based on simultaneous emission and detection of multiple photons (photon-antibunching) under realistic experimental conditions. In theory, it should be possible to estimate the number of molecules with errors of less than 1% by using novel photo-stabilizing agents. The proposed method puts into perspective its application for high-resolution microscopy without the need for photo-switching or photo-activation of fluorescence dyes.     

A Feasible Add-On Upgrade on a Commercial Two-Photon FLIM Microscope for Optimal FLIM-FRET Imaging of CFP-YFP Pairs

Fluorescence lifetime imaging microscopy (FLIM) based on time-correlated single photon counting (TCSPC) is a widely used method for fluorescence resonance energy transfer (FRET). Here we report a feasible add-on approach to upgrade a commercial two-photon FLIM microscope into a single-photon FLIM microscope which provides optimal FLIM-FRET imaging of FRET pairs consisting of cyan fluorescent proteins (CFPs) as the donor and yellow fluorescent proteins (YFPs) as the acceptor. The capability of the upgraded system is evaluated and discussed, and the imaging performance of the system is demonstrated using FLIM-FRET experiments with a representative CFP-YFP FRET pair (mCerulean-mCitrine).     

基于SiPM和TCMPC的时间分辨拉曼散射测量技术研究

报道了一种基于硅光电信增管(SiPM)的时间相关多光子计数(TCMPC)技术并将其应用于时间分辨拉曼散射测量。相比于常规基于光电倍增管(PMT)或单光子雪崩二极管(SPAD)的时间相关单光子(TCSPC)技术,由于SiPM可以分辨信号脉冲的具体光子数,基于SiPM的TCMPC技术消除了信号脉冲包含的光子数必须小于等于1的限制,光子计数效率提高了10倍以上,大大节省了测量时间。此外,多光子测量比单光子测量能够得到更好的时间分辨率,时间分辨拉曼散射系统的仪器响应函数(IRF)从单光子81.4 ps缩短至双光子59.7 ps,因而可以用更窄的时间门限抑制荧光本底等噪声对拉曼散射测量的影响。使用TCMPC技术测量CCl₄在0.5和1.5 p.e.两个不同光子数阈值的拉曼峰的峰本比,后者较高的光子数阈值能进一步降低SiPM暗计数噪声的影响,增加了拉曼信号测量的信噪比,测量得到的CCl₄ 459 cm-1拉曼峰的峰本比是前者的6.4倍。将所述新的拉曼散射测量技术与基于PMT和锁相放大器(LIA)的传统拉曼散射测量技术进行了比较研究,前者由于可以使用仅有数十皮秒的测量门限,可以有效抑制荧光、环境杂散光和SiPM暗计数等噪声的影响,所得光谱具有更好的峰本比,测得CCl₄的459 cm-1拉曼峰和Si的一阶拉曼峰的峰本比分别是后者的3.9倍和5.5倍。     

基于时间相关单光子计数技术的密码芯片光辐射分析

密码芯片运行时的光辐射可泄露其操作和数据的重要特征信息. 基于单光子探测技术, 设计并构建了针对CMOS半导体集成电路芯片光辐射信号的采集、传输、处理和分析的光电实验系统. 以AT89C52单片机作为实验对象, 采用时间相关单光子计数技术, 对不同工作电压下密码芯片的光辐射强度进行了对比, 分析了芯片指令级光辐射信息的操作依赖性和数据依赖性. 此外, 使用示波器对时间相关单光子计数技术在芯片光辐射分析上的可行性进行了验证. 实验结果表明, 采用时间相关单光子计数技术对密码芯片进行光辐射分析, 是一种直接有效的中低等代价光旁路分析攻击手段, 对密码芯片的安全构成了严重的现实威胁.     

Subcentimeter depth resolution using a single-photon counting time-of-flight laser ranging system at 1550 nm wavelength

We demonstrate subcentimeter depth profiling at a stand off distance of 330 m using a time-of-flight approach based on time-correlated single-photon counting. For the first time to our knowledge, the photon-counting time-of-flight technique was demonstrated at a wavelength of 1550 nm using a superconducting nanowire single-photon detector. The performance achieved suggests that a system using superconducting detectors has the potential for low-light-level and eye-safe operation. The system's instrumental response was 70 ps full width at half-maximum, which meant that 1 cm surface-to-surface resolution could be achieved by locating the centroids of each return signal. A depth resolution of 4 mm was achieved by employing an optimized signal-processing algorithm based on a reversible jump Markov chain Monte Carlo method.     

A method for on-line background subtraction in frequency domain fluorometry

Time-resolved fluorescence determinations on biochemical samples are often complicated by contributions from background. In the time-correlated single-photon counting method, background subtraction is a routine procedure. A limitation of frequency domain fluorometry, however, arises from the difficulty of performing this operation. This limitation has become increasingly significant as frequency domain methods are being applied to evermore complex biological systems using the higher-frequency capabilities of modern instrumentation. We have devised a method for such a correction in the frequency domain, regardless of the complexity of the background decay, based on measurement of the background phasor and subsequent subtraction from the sample phasor. This method is applicable to both lifetime and dynamic polarization measurements, and it can be readily implemented on commercially available frequency domain fluorometers. Decay curves may be accurately recovered from samples containing background contributions ranging from less than 5% to greater than 90% of the total signal intensity.     

Single-Photon Counting in Luminescent Analysis with Micro- and Millisecond Decay Times

Optics and Spectroscopy - Main methods of photon counting, as well as particularities in detecting micro- and millisecond decay times using a single-photon gated avalanche photodiode, are...     

皮秒光脉冲在光纤中的传播特性测量

文章讨论了皮秒激光脉冲在单模光纤传输中光纤介质的色散及非线性折射率等效应对皮秒光信号的影响，利用时间相关单光子计数系统，观测了皮秒激光脉冲在单模光纤中的展宽及皮秒自聚焦现象，并对光纤中皮秒光脉冲的畸变进行了检测与研究。     

Binding of Biotin to Streptavidin: A combined fluorescence correlation spectroscopy and time-resolved fluorescence study

The Biotin-Streptavidin complex is a widely studied system in biology and biophysics, because of its extremely strong non-covalent binding affinity. The latter is often exploited to link molecules to substrates or to one another. However, the details of the Biotin-Streptavidin binding have not been fully elucidated so far. Particularly, the role of cooperative effects in enhancing the binding affinity has not been clarified. Our long-term aim is to investigate this point by implementing two complementary approaches, fluorescence correlation spectroscopy and time-correlated single-photon counting. As both methods rely on the analysis of fluorescence signals, biotin labeled with Atto-550-dye was used. In this work, in order to get a first overview of the system, we analyzed solutions in three paradigmatic ranges of Biotin-to-Streptavidin concentration ratio. Fluorescence correlation spectroscopy measurements allowed us to extract diffusion times of free biotin and of biotin-Streptavidin complexes, and also to gain information about the dynamics of the intersystem crossing between the first excited triplet and the first excited singlet states. Time-correlated single-photon counting made it possible to derive the lifetimes of the different species in solution, as well as to deduce relevant information about the relative abundance of Streptavidin-complexed and free Biotin.     

荧光衰减曲线的一种多指数拟合方法

在荧光衰减测量中，总要遇到荧光衰减解卷积，进行非线性参数拟合问题。目前已有的非线性求极小值方法，都对参数估计值非常敏感，而且计算程序运行时间较长。我们建立了一种基于经典的步长加速法实现非线性最小二乘拟合，进行荧光衰减解卷积的方法，按程序给出参数初值，即能解卷积。     

Lifetime measurement of the 9s level of atomic francium

We use two-photon resonant excitation and time-correlated single-photon counting techniques on a sample of 210Fr atoms confined and cooled in a magneto-optical trap to measure the lifetime of the 9s excited level. Direct measurement of the decay through the 7P(3/2) level at 851 nm yields a lifetime of 107.53 +/- 0.80 ns.     

Fluorescence methods to study lipid-protein association: The interaction of protein kinase C with lipid-loaded mixed micelles

The interaction of protein kinase C with lipids was studied in a mixed micellar system. Two fluorescence spectroscopic methods are presented with a different but complementary information content. Diffusion monitored by fluorescence correlation spectroscopy provides information on the interaction of the protein with the whole lipid aggregate. Resonance energy transfer from tryptophans to pyrene-labeled lipids monitored by time-correlated single-photon counting supplies information on the interaction of the protein with specific lipid cofactors within the micelle. The results can be extended to postulate new mechanisms for the activation of protein kinase C by the signal transduction cascades in the cell. Both fluorescence spectroscopic methods can be easily applied to other protein systems which interact with lipids.     

Hyper Rayleigh Scattering Yields Improved Response Function in Analysing 2-Photon Excited Fluorescence

An accurate instrumental response function is needed to conclusively deconvolute fluorescence data based on time-correlated single-photon counting (TCSPC) and multiphoton excitation. Routinely the response function is measured as Rayleigh scattering (RS) from a colloidal solution, even if the excitation is a multiphoton event. Present work demonstrates that a response function obtained as hyper Rayleigh scattering (HRS) provides a better choice for deconvolution of 2-photon excited fluorescence decays. The 1- and 2-photon response functions were monitored as RS and HRS from colloidal gold particles at 800 and 400 nm, respectively.     

Multiplexed time-correlated single-photon counting

We review the technique of multiplexed time-correlated single-photon counting whereby multiple fluorescence decay curves are recorded in parallel by statistically time-sharing a single time-to-amplitude converter. Application of the multiplexing technique to measuring the fluorescence lifetime topography of a self-absorbing sample is demonstrated. Further possibilities are discussed for multiplexed optical fiber sensor networks with built-in intelligence for detecting and discriminating between different metal ions in solution.     

Estimate of etched tracks by optical method and spark counting

Solid State Nuclear Track Detectors (SSNTD) are commonly used for long term measurements of radon, thoron and progeny concentrations. In the present study, we compared the optical counting method and the spark counting technique for counting the alpha tracks on LR 115 track detector films. The paper discusses the various parameters that are innate in the process. More than 300 films were counted by both optical microscope and spark counter and the results are compared. The overall results show that the tracks obtained by spark counting are marginally less compared to the optical measurement. A linear fit of the data gives a slope less that one, which indicates that both the methods are almost in good agreement for counting the tracks when the track density is low. At higher track densities the spark counter gives an underestimation with respect to actual number of tracks formed, which could be corrected using a relation obtained between the tracks measured by optical method and spark counting.      

Broadband optical limiting performance of polymer-wrapped carbon nanotubes in the orange-NIR region

Soluble multi-walled carbon nanotubes (MWNTs) have been obtained by noncovalent modification with poly [2-methoxy,5-(2′-ethyl-hexyloxy)-p-phenylene vinylene] (MEH-PPV). For the composite MWNT/MEH-PPV, there is π-π interaction between the MEH-PPV and MWNTs in addition to the wrapping of the polymer. The nonlinear optical transmittance was measured using a nanosecond optical parametric oscillator pumped with a Nd:YAG system. Excellent optical limiting performance of the composite MWNT/MEH-PPV was observed both in the visible region of 590-680 nm and at the wavelength of 1064 nm. By means of time-correlated single-photon counting fluorescence measurement, an explanation based on the nonlinear absorption of MWNT dominated by the intermolecular energy transfer was proposed.     

硒化锌单晶薄膜近带边发射的荧光寿命

在2K下,对常压MOCVD方法生长的ZnSe单晶薄膜进行了光致荧光近带边发射谱的测量和辨认.并采用激光超短脉冲技术和时间相关单光子计数技术相结合的方法,对其在2K下近带边发射的荧光寿命进行了测量和分析,并讨论了自由激子弛豫过程.