Picoharp 300

Manufactured by PicoQuant

Sourced in Germany

The PicoHarp 300 is a time-correlated single-photon counting (TCSPC) module for time-resolved measurements. It provides high-resolution time-tagging of detected photons with a time resolution of up to 1 picosecond. The PicoHarp 300 is capable of recording photon arrival times with respect to a reference signal, enabling time-resolved fluorescence and phosphorescence measurements.

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Lab products found in correlation

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Close spelling variants of this product

PicoHarp 300 TCSPC system (4 citations) FluoTime 200/PicoHarp 300 spectrometer (2 citations) PicoHarp 300 TCSPC Module (10 citations) PicoHarp (3 citations)

93 protocols using picoharp 300

Time-Resolved Photoluminescence of Perovskites

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TRPL of MAPbBr₃, MAPbI₃, and FAPbBr₃ was measured by a home-built setup. A 405 nm pulsed femtosecond laser (Mai Tai) with a repetition rate of 4 MHz was focused onto the surface of the sample using an objective lens (Olympus, 20×, SApo, NA = 0.7), generating a Gaussian-distribution spot with a FWHM of 1366 nm (for FAPbBr₃, the repetition rate was 0.5 MHz). All measurements were done with an average incident power of ≈40 nW. Then, the PL from the photoexcitation, together with the reflected and scattered excitation signal, was collected by the same objective lens. The resulting collimated beam passed through a long-pass filter with a cutoff at λ = 425 nm to remove the reflected and scattered excitation light. The resulting PL signal was further collected through a multimode fiber (Thorlabs, 50 μm in diameter, 5 m) with a pair of coupler and collimator (Thorlabs RC08FC-P01) and sent to an APD (MPD PDM Series 20 μm), which was attached to the timing electronics (PicoQuant PicoHarp 300).
The TRPL of MAPbCl₃ was measured by a home-built setup. 515 nm femtosecond laser was used to excite PL signal by two-photon absorption. A 450 nm short pass filter was used to remove the reflected and scattered excitation light. The resulting PL signal was further collected and sent to an APD (MPD PDM Series 20 μm) which is attached to the timing electronics (PicoQuant PicoHarp 300).

Chen X.G., Lin L., Huang G.Y., Chen X.M., Li X.Z., Zhou Y.K., Zou Y., Fu T., Li P., Li Z, & Sun H.B. (2024). Optofluidic crystallithography for directed growth of single-crystalline halide perovskites. Nature Communications, 15, 3677.

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Picosecond Laser FLIM Imaging of o-BMVC

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The setup of the FLIM system consisted of a picosecond diode laser (laser power, 5 mW) with an emission wavelength of 470 nm (LDH470; PicoQuant, Germany) and a ~ 70 ps pulse width for the excitation of o-BMVC under a scanning microscope (IX-71 and FV-300; Olympus, Japan). The fluorescent signal from o-BMVC was collected using a 60 × NA = 1.42 oil-immersion objective (PlanApoN; Olympus, Japan), passed through a 550/88 nm bandpass filter (Semrock, USA), and detected using a single-photon avalanche diode (SPAD) (PD-100-CTC; Micro Photon Devices, Italy). The fluorescence lifetime was recorded and analyzed using a time-correlated single-photon counting (TCSPC) module and software (PicoHarp 300 and SymPhoTime v5.3.2; PicoQuant, Germany). FLIM images were constructed from pixel-by-pixel lifetime information.

Tseng T.Y., Shih S.R., Wang C.P., Lin S.J., Jan I.S., Wang C.L., Liu S.Y., Chang C.C., Lou P.J, & Chang T.C. (2021). Pilot imaging study of o-BMVC foci for discrimination of indeterminate cytology in diagnosing fine-needle aspiration of thyroid nodules. Scientific Reports, 11, 23475.

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Intracellular Doxorubicin Imaging by FLIM

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Each imaging experiment started 3 h after the incubation of the cells with GO-DOX and Doxoves^®, without washing the cells to remove the excess of non-internalized compounds. For imaging, a Leica SP5 confocal microscope (Leica Microsystems AG, Wetzlar, Germany) was used, equipped with an Argon laser for excitation at 488 nm. Doxorubicin emission was collected in the 500-650 nm range. All experiments were performed by using the same imaging settings (e.g., magnification, laser power, etc.). The quantification of the recorded signal was carried out by Image J software (National Institutes of Health, Bethesda, Maryland, USA). During the experiment, the cells were maintained in a thermostatic chamber (37 °C, 5% CO₂). For FLIM experiments, all analyzed samples were excited at 470 nm with a pulsed diode laser operating at 40 MHz (average power: 10-20 μW at the sample) and collecting the emission in the 500-650 nm range by a photomultiplier tube interfaced with a time-correlated single photon counting card and setup (PicoHarp 300, PicoQuant, Berlin). Phasor analysis of lifetime data was performed by SimFCS software (www.lfd.uci.edu, University of California at Irvine).

Quagliarini E., Di Santo R., Pozzi D., Tentori P., Cardarelli F, & Caracciolo G. (2020). Mechanistic Insights into the Release of Doxorubicin from Graphene Oxide in Cancer Cells. Nanomaterials, 10(8), 1482.

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Porphyrin Photophysics via Confocal FLIM

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Standard porphyrins PPIX and CPI were either analyzed in methanol solution (about 1 × 10⁻⁵ M) or crystallized from solution on a glass coverslip by slow evaporation of the solvent. The measurements were performed using a Leica TCS SP5 inverted confocal microscope (Leica Microsystems, Wetzlar, Germany). An external pulsed diode laser provided excitation at 405 nm and 640 nm, while a TCSPC acquisition card (PicoHarp 300, PicoQuant, Berlin, Germany) connected to internal spectral photomultipliers allowed for detection. On the basis on the lifetime values, laser repetition rate was fixed at 20 or 40 Hz. Image size was set to 256 × 256 pixels and scan speed was modulated between 200 and 400 Hz (lines per second). The detection wavelength range was set between 580 and 720 nm for λ_ex= 405 nm and between 650 and 750 nm for λ_ex= 640 nm thanks to the built-in AOBS detection system. About 200–300 photons per pixel were collected for each measurement, at a photon counting rate of 100–200 kHz. Data collected from different ROIs in the biofilms images were averaged. Globals for Images—SimFCS v.2 (Globals Software by LFD-UCI, Irvine, CA, USA, available at www.lfd.uci.edu, accessed on 1 September 2021) was used to calculate phasors from the FLIM images.

Battisti A., Morici P, & Sgarbossa A. (2021). Fluorescence Lifetime Imaging Microscopy of Porphyrins in Helicobacter pylori Biofilms. Pharmaceutics, 13(10), 1674.

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Ultrafast Perovskite Optoelectronic Characterization

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Bulk MAPbI₃, MAPbBr₃, and CsPbBr₃ films were measured using an yttrium-aluminum-garnet–Nd laser (Spitlight Compact 100) emitting at 355 or 532 nm with a pulse length of ~10 ns and an energy of 50 μJ cm⁻² in both cases. The spot size was 5 mm in diameter. Signal detection was performed with a Shamrock spectrometer (SR-303i-A), equipped with an intensified charge-coupled device camera [Andor iStar DH320T-18U-73 (gate step, 2.5 ns; gate width, 2.5 ns)]. The samples were kept under vacuum during the measurement. Corresponding shorter-lived perovskite nanocrystals in 5–mA cm⁻² npSi templates were investigated using a superconducting single photon detector (SSPD; Scontel Superconducting Nanotechnology) together with a time-correlated single-photon counting system (PicoHarp 300 by PicoQuant). The time resolution of the SSPD system is about 300 ps. Samples were excited by a pulsed diode laser (405 nm) delivering ~1-ns pulses (FWHM) at repetition rates of 1 MHz and pulse energies of approximately 5 pJ. A microscope objective was used to focus the laser onto a spot with a 10-μm diameter on the sample surface. The sample emission was collected using the same microscope objective, and the photons were guided through a single-mode fiber to the SSPD. A long-pass filter was applied to block the excitation before entering the fiber connected to the SSPD.

Demchyshyn S., Roemer J.M., Groiß H., Heilbrunner H., Ulbricht C., Apaydin D., Böhm A., Rütt U., Bertram F., Hesser G., Scharber M.C., Sariciftci N.S., Nickel B., Bauer S., Głowacki E.D, & Kaltenbrunner M. (2017). Confining metal-halide perovskites in nanoporous thin films. Science Advances, 3(8), e1700738.

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Simultaneous DCS and SCOS Acquisition

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We use a DCS system consisting of a single channel SPAD (SPCM-AQ4C, Excelitas) with a time tagger (PicoHarp 300, PicoQuant). The source fiber and CW laser are shared with the fiber-based SCOS system. The DCS system is synchronized with the fiber-based SCOS for simultaneous data acquisition. The intensity temporal autocorrelation function (

g_{2} (τ)

) was generated from the raw intensity counts for time delay from 1 µs to 14.3 ms, with exponential increase in step between time delays. The bin size was set at 1 µs for the measurement time of 100 ms. The laser coherence (β) was estimated from fitting the

g_{2} (τ)

curve averaged over the duration of the measurement to an exponential decay function given by^{36 (link),37 (link)}:

g_{2} (τ) = 1 + β \exp (\frac{- 2 τ}{τ_{c}})

where τ is the time delay, and τ_c is the decorrelation time. For each g₂(τ) curve averaged over the measurement time the same exponential decay function and prior estimate of laser coherence is used to estimate the decorrelation time. The least-square curve fit was done using MATLAB’s lsqcurvefit function with implementation of the Levenberg-Marquardt algorithm.

Kim B., Zilpelwar S., Sie E.J., Marsili F., Zimmermann B., Boas D.A, & Cheng X. (2023). Measuring human cerebral blood flow and brain function with fiber-based speckle contrast optical spectroscopy system. Communications Biology, 6, 844.

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Photoluminescence Lifetime Measurement

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When obtaining the PL intensity time traces and lifetimes, photons were detected using single photon counting APDs (SPCM-AQ4C, Perkin Elmer). TTTR T3 mode (1 ns resolution) is used for the lifetime measurements with using two channels of the PicoHarp 300 (PicoQuant). Both the T2 and T3 mode are used for the PL intensity time trace. All collected data were reconstructed and analyzed using algorithms, coded utilizing the MATLAB software, and fitted employing the ORIGIN 2016 software. The temporal resolution of the photon detection system used in this study was estimated to be approximately 0.65 ns by measuring temporally short laser pulses (~3 ps), and de-convoluting the measured curves.

Trinh C.T., Minh D.N., Ahn K.J., Kang Y, & Lee K.G. (2020). Verification of Type-A and Type-B-HC Blinking Mechanisms of Organic–Inorganic Formamidinium Lead Halide Perovskite Quantum Dots by FLID Measurements. Scientific Reports, 10, 2172.

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Optical Characterization of Single-Photon Emitters

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The optical properties of the single-photon emission were measured using a home-built confocal microscope setup consisting of a system of two sets of galvanometers to control the excitation and emission individually. Pulsed (SuperK EXTREME (NKT Photonics, Birkeød, Denmark)) or continuous-wave (CW) lasers (OBIS 532 nm LS (Coherent Inc., Santa Clara, CA, USA)) with a wavelength of 532 nm were used as the pumping source. Emission from the h-BN was collected by a 100× objective lens that had a numerical aperture of 0.90 and was coupled to the single-mode fiber to send the PL to either the Avalanche photodiode (APD) or the spectrometer (Acton SP2500 (Teledyne Princeton Instruments, Trenton, NJ, USA)). To measure the second-order correlation, we used a Hanbury Brown and Twiss (HBT) interferometer setup composed of two APDs and a time-correlated single-photon counting module (Picoharp 300 (PicoQuant, Berlin, Germany)). The emission from the sample was divided into two APDs using a 50:50 fiber beam splitter. Polarization measurements were conducted by placing a linear polarizer in front of the fiber coupler.

Jeong K.Y., Lee S.W., Choi J.H., So J.P, & Park H.G. (2020). Integration of Single-Photon Emitters in 2D Materials with Plasmonic Waveguides at Room Temperature. Nanomaterials, 10(9), 1663.

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Time-Resolved Fluorescence of NADH-Enzyme Complexes

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In this system, the excitation pulse centered at 340 nm (with 10 MHz repetition) was generated by a picosecond pulsed diode laser (PDL 800-B, PicoQuant). The fluorescence was recorded using a time-correlated single photon counting module (PicoHarp 300, PicoQuant) and a single photon counting PMT (PMA165A-N-M, PicoQuant). The instrument response function was found to be ~440 ps in width by measuring the Rayleigh scattering of excitation pulses in a suspension of 0.34 wt% SiO₂ nanoparticles in water. More details about the system can be found elsewhere.³⁴ For constructing decay associated spectra (DAS), the fluorescence was scanned (in the range 420–560 nm for NADH–MDH and 420–500 nm for NADH–LDH) with 10 nm bandwidth and analyzed by a global fitting technique³⁵ using a biexponential model. The fits in this work usually yielded x² values of 1.01–1.15.
Steady-state absorption spectra were measured with a UV–visible spectrophotometer (TU1901, Beijing Purkinje General Instrument Co. Ltd.). Steady-state fluorescence spectra were recorded using a commercial spectrofluorometer (FluoroMax-4, Horiba). The decay associated spectra were normalized to the steady-state spectra of NADH–MDH/LDH in Tris–HCl buffers.

Cao S., Li H., Liu Y., Wang M., Zhang M., Zhang S., Chen J., Xu J., Knutson J.R, & Brand L. (2020). Dehydrogenase Binding Sites Abolish the “Dark” Fraction of NADH: Implication for Metabolic Sensing via FLIM. The journal of physical chemistry. B, 124(31), 6721-6727.

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Time-resolved Photoluminescence of CdS Photoanodes

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Emission lifetimes of the CdS containing photoanodes immersed in aqueous electrolyte (1 M HCOONa) were acquired with a Picoquant Picoharp 300 time correlated single photon counting at a 4 ps resolution by using a 480 nm pulsed LED source. Levenberg-Marquardt fitting/deconvolution of the decay histogram was accomplished with a tri-exponential function by the dedicated Fluofit program. In general, fits satisfied the statistical acceptability criteria, with χ2 ≈1 and residuals R(i) = W(i)(Decay(i) − Fit(i)) < 4 standard deviations fluctuating around 0 within all the fitting intervals. In the R(i) formula W(i) = 1/(decay(i))1/2 defines the intensity weight in a given channel (i) according to the Poisson distribution while Decay(i) and Fit(i) are the experimentally measured and calculated decay values respectively.

Mazzanti M., Milani M., Cristino V., Boaretto R., Molinari A, & Caramori S. (2022). Visible Light Reductive Photocatalysis of Azo-Dyes with n–n Junctions Based on Chemically Deposited CdS. Molecules, 27(9), 2924.

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