Ldh d c 640

Manufactured by PicoQuant

Sourced in Germany

The LDH-D-C-640 is a picosecond diode laser from PicoQuant. It is a solid-state device that can generate pulsed light at a wavelength of 640 nanometers. The laser is designed for use in various scientific and research applications.

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Lab products found in correlation

Spcm cd3516h, by Excelitas (6 mentions) Microtime 200 fluorescence lifetime microscope, by PicoQuant (4 mentions) Ldh d ta 530, by PicoQuant (4 mentions) Uplansapo 60x 1.2w, by Olympus (3 mentions) Hydraharp 400, by PicoQuant (3 mentions) Fv1200, by Olympus (3 mentions) P 733.2cd, by Physik Instrumente (3 mentions) Tau spad, by PicoQuant (2 mentions) Uplsapo 100xo, by Olympus (2 mentions) Labview 2009, by National Instruments (2 mentions) P 517.3cl, by Physik Instrumente (2 mentions) Razoredge lp 647, by IDEX Corporation (2 mentions) τ spad 100, by PicoQuant (2 mentions) Uplsapo 60xo, by Olympus (2 mentions) Spc 830, by Becker & Hickl (2 mentions) Brightline, by IDEX Corporation (1 mentions) Objective len, by Olympus (1 mentions) Spcm aqr 14, by PerkinElmer (1 mentions) Sepia 2, by PicoQuant (1 mentions) Uplansapo, by Olympus (1 mentions)

14 protocols using ldh d c 640

Confocal FCS Measurements with SUVs

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FCS measurements were performed on
a commercial, epi-illuminated, confocal laser scanning microscope
(Olympus FV1200). Solution samples with SCy7 fluorophores, as free
labels, or in SUV preparations (as described above) were excited by
the focused beam of a 638 nm (338 nm, 1/e² radius) or 780
nm diode laser (LDH-D-C-640 and LDH-D-C-780, both from PicoQuant GmbH,
Berlin) in continuous wave. The emitted fluorescence was collected
back through the microscope objective (UPlanSApo 60x/1.2w, Olympus),
passed through a dichroic mirror (ZT405/488/635rpc-UF2, Chroma or
T800lpxr-xt-UF2, Chroma) and an emission filter (HQ720/150, Chroma,
809/81 Brightline, Semrock, Semrock or 835/70 Brightline, Semrock),
and focused onto a pinhole (50 μm diameter) in the back focal
plane. The fluorescence signal was finally split and directed on two
avalanche photodiodes (Tau-SPAD, PicoQuant GmbH, Berlin), whose signals
were collected by a data acquisition card (Hydraharp 400, Picoquant,
Berlin).

Sandberg E., Piguet J., Kostiv U., Baryshnikov G., Liu H, & Widengren J. (2023). Photoisomerization of Heptamethine Cyanine Dyes Results in Red-Emissive Species: Implications for Near-IR, Single-Molecule, and Super-Resolution Fluorescence Spectroscopy and Imaging. The Journal of Physical Chemistry. B, 127(14), 3208-3222.

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Confocal FCS of Cy5 in Aqueous Solution

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FCS measurements were performed on a commercial, epi-illuminated, confocal laser scanning microscope (Olympus, Tokyo, Japan, FV1200). Cy5 in aqueous solution was excited with a focused beam (338 nm, 1/e² radius) of a 638 nm laser (LDH-D-C-640 from PicoQuant GmbH, Berlin, Germany) in continuous wave. The emitted fluorescence was collected back through the microscope objective (UPlanSApo 60x/1.2 w, Olympus, Tokyo, Japan), passed through a dichroic mirror (ZT405/488/635rpc-UF2, Chroma), an emission filter (HQ720/150, Chroma, 680 nm blocking edge Brightline, Semrock, in combination with last mentioned filter or 710/40 Brightline, Semrock), and focused onto a pinhole (50 µm diameter) in the back focal plane. The fluorescence signal was finally split and directed on two avalanche photodiodes (Tau-SPAD, PicoQuant GmbH, Berlin, Germany), whose signals were collected with a data acquisition card (Hydraharp 400, Picoquant, Berlin, Germany).

Sandberg E., Piguet J., Liu H, & Widengren J. (2023). Combined Fluorescence Fluctuation and Spectrofluorometric Measurements Reveal a Red-Shifted, Near-IR Emissive Photo-Isomerized Form of Cyanine 5. International Journal of Molecular Sciences, 24(3), 1990.

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Single-molecule FRET of SthK Channels

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After the preparation of immobilized, fluorophore-labeled SthK nanodiscs on the slide surface, sm-FRET data was acquired from individual SthK channels using a MicroTime 200 Fluorescence Lifetime Microscope from PicoQuant. A donor excitation laser (532 nm; LDH-D-TA-530; Picoquant, Berlin, Germany) and acceptor excitation laser (637 nm; LDH-D-C-640; Picoquant) were used with a Pulsed Interleaved Excitation (PIE) scheme to excite the fluorophores. Emitted photons were collected back through the objective lens (100×1.4 numerical aperture; Olympus). Emission filters for the donor (550 nm; FF01-582/64; AHF, Tübingen-Pfrondorf, Germany or Semrock, Rochester, NY) and acceptor (650 nm 2XH690/70; AHF) were used to select photons for each detection channel. Photons were then passed to two SPAD photodiodes (SPCM CD3516H, Excelitas technologies, Waltham, MA) to determine the fluorescence intensity for each fluorophore. The donor and acceptor fluorescence intensities over time were recorded for one SthK channel at a time and later analyzed as described below.

Gao X., Schmidpeter P.A., Berka V., Durham R.J., Fan C., Jayaraman V, & Nimigean C.M. (2022). Gating intermediates reveal inhibitory role of the voltage sensor in a cyclic nucleotide-modulated ion channel. Nature Communications, 13, 6919.

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Single-molecule Fluorescence Imaging Setup

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Single-molecule fluorescence transients are measured with a custom built confocal setup based on an inverted microscope (IX71, Olympus) with a high NA oil immersion objective (60X, NA 1.35, UPLSAPO 60XO for rectangular DNA origami; 100x/NA 1.40, UPLSAPO100XO for DNA nanopillar and ZMWs, both Olympus). The ATTO647N dye molecules are excited at 640 nm with an 80 MHz pulsed laser diode (LDH-D-C-640, Picoquant, 2.5 μW for rectangular DNA origami, 1.5 μW for DNA nanopillar measurements, 5 μW for ZMW measurements). A combination of linear polarizer and quarter wave plate ensures circular polarization of the laser beam which is guided to the sample by a dichroic beamsplitter (Dualband z532/633, AHF).The surface with immobilized DNA origamis is imaged by scanning the sample with a piezo stage (P-517.3CL, Physik Instrumente). From that image, molecules are selected and placed in the laser focus for time resolved analysis. The resulting fluorescence is collected by the same objective and separated from the excitation light after focusing on a 50 μm pinhole by two filters (Bandpass ET 700/75m, AHF; RazorEdge LP 647, Semrock). The signal is detected by a single photon counting module (τ-SPAD 100, Picoquant) and a PC-card for time-correlated single-photon counting (SPC-830, Becker&Hickl). The raw data is processed with custom made software (LabView2009, National Instruments).

Holzmeister P., Pibiri E., Schmied J.J., Sen T., Acuna G.P, & Tinnefeld P. (2014). Quantum yield and excitation rate of single molecules close to metallic nanostructures. Nature communications, 5, 5356.

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Single-Molecule FRET Microscopy Protocol

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smFRET measurements were collected using a custom-built PicoQuant Microtime 200 Fluorescence Lifetime Microscope. To characterize the fluorescent behavior of both donor and acceptor fluorophores as well as the efficiency of the energy transfer between donors and acceptors, acquisition was conducted using both 532 nm (LDH-D-TA-530; Picoquant, Berlin, Germany) and 637 nm (LDH-D-C-640; Picoquant) lasers using pulsed interleaved excitation at 80 MHz. During scanning, the slide was immobilized on a scanning x-y-z piezo stage (P-733.2CD; Physik Instrumente) and observed through a 100× oil-immersed lens (100 × 1.4 NA; Olympus, Tokyo, Japan). Photons from samples post-excitation were then collected back through the objective and separated through a dual band dichroic beam splitter (Zt532/640rpc-UF3; AHF/Chroma, Bellows Falls, VT, USA). Prior to detection, photons were then filtered through emission filters (550 nm (FF01-582/64; AHF/Semrock, Rochester, NY, USA) for the donor or 650 nm (2XH690/70; AHF, Tübingen-Pfrondorf, Germany) for the acceptor) and into two SPAD photodiodes (SPCM CD3516H; Excelitas Technologies, Waltham, MA, USA).

Gonzalez C.U., Carrillo E., Berka V, & Jayaraman V. (2021). Structural Arrangement Produced by Concanavalin A Binding to Homomeric GluK2 Receptors. Membranes, 11(8), 613.

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Confocal FCS Measurements and Analysis

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FCS measurements were
performed on a commercial, epi-illuminated, confocal laser scanning
microscope (Olympus FV1200), with the samples excited by the focused
beam of a 640 nm diode laser (LDH-D-C-640, PicoQuant GmbH, Berlin)
and the emitted fluorescence collected through the same microscope
objective (UPlanSApo 60x/1.2w, Olympus). The normalized
autocorrelation of the recorded fluorescence intensity fluctuations
(the FCS curves) typically displayed relaxation terms attributed to
diffusion (G(τ)) and fluorophore relaxation
into a dark transient state (G_T(τ))
and were analyzed as previously described,^{13 (link),14 (link)} using a Levenberg-Marquart nonlinear least-squares optimization,
with no weighting on the residuals. See Supplementary, Section S6 for further details.

Sandberg E., Demirbay B., Kulkarni A., Liu H., Piguet J, & Widengren J. (2023). Fluorescence Bar-Coding and Flowmetry Based on Dark State Transitions in Fluorescence Emitters. The Journal of Physical Chemistry. B, 128(1), 125-136.

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Single-Molecule Fluorescence Microscopy for PET-FCS

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PET-FCS measurements were performed on a custom-built confocal single-molecule fluorescence microscope. The sample was excited with a diode laser (LDH-D-C-640, PicoQuant) operated in continuous wave mode at an average laser power of 280 μW as measured before the aperture of the objective. The fluorescence signal was passed through a pinhole and split on two avalanche photodiodes (SPCM-AQR-14, Perkin Elmer Optoelectronics) by a 50:50 beam splitter to avoid detector dead time, connected to two independent channels of the time-correlated single photon counting (TCSPC) hardware (HydraHarp400, PicoQuant). Fluorescence signal was passed through an emission filter (ET670/30, AHF Analysentechnik). The range of the emission filter was chosen as to avoid detector crosstalk due to the breakdown flash of the APDs [57 ]. Individual photon arrival times were recorded with 16 ps resolution. S-peptide was dissolved in standard PBS buffer with 0.005% Tween-20 to prevent sticking to the cover slide surface, and diluted to a final concentration of ∼1 nM. FCS data was collected over a time of 16 h at room temperature.

Luitz M.P., Barth A., Crevenna A.H., Bomblies R., Lamb D.C, & Zacharias M. (2017). Covalent dye attachment influences the dynamics and conformational properties of flexible peptides. PLoS ONE, 12(5), e0177139.

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Single Molecule FRET Microscopy

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Single molecule FRET measurements were acquired using a custom-built PicoQuant MicroTime 200 Fluorescence Lifetime Microscope. smFRET data acquisitions were conducted using pulsed interleaved excitation at 80 MHz. Both 532 nm (LDH-D-TA-530; Picoquant) and 637 nm (LDH-D-C-640; Picoquant) lasers were simultaneously used to characterize the fluorescent behavior of both fluorophores and the efficiency of energy transfer between molecules potentially showing FRET. The sample slide was immobilized on a scanning x-y-z piezo stage (P-733.2CD; Physik Instrumente) while being excited and observed through a 100x oil immersed lens (100 × 1.4 NA; Olympus). The photons emitted from the sample post-excitation were collected back through the objective, separated through a dual band dichroic beam splitter (Zt532/640rpc-UF3; AHF/Chroma) and sent to two SPAD photodiodes (SPCM CD3516H; Excelitas technologies) preceded by excitation filters. A 550 nm (FF01-582/64;AHF/Semrock) and 650 nm (2XH690/70;AHF) emission filter were used for the donor and acceptor channels, respectively. All acquisitions were performed in the presence of a photo-stabilizer and oxygen scavenging solution buffer system (ROXS).

Litwin D.B., Carrillo E., Shaikh S.A., Berka V, & Jayaraman V. (2019). The structural arrangement at intersubunit interfaces in homomeric kainate receptors. Scientific Reports, 9, 6969.

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Dual-Laser Confocal Fluorescence Fluctuation Spectroscopy

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Measurements were performed on an Olympus IX73 inverted microscope stand equipped with a 1.2 N.A. water-immersion 60× superapochromat objective (UplanSApo; Olympus) and suitable emission and excitation bandpass filters (Semrock and AHF). Two pulsed diode lasers (LDH-P-FA-530 and LDH-D-C-640; PicoQuant) were operated at 40 MHz for pulse interleaved excitation dcFLCCS (Sepia II; PicoQuant). Emitted photons were detected in two separated channels coupled with two SPAD detectors (SPCM CD3516H; Excelitas) and a time-correlated single-photon counting unit to generate picosecond histograms also called lifetime spectra (16-ps resolution; HydraHarp 400) from the statistical photon arrival times. The laser powers were set to 20 µW for the LDH-P-FA-530 and to 17 µW for the LDH-D-C-640 laser and the intensity fluctuation recorded for 120 s with a correlation integration time taken as 2 s. The confocal volume was calibrated using free dyes of known diffusion constants D (using Rhodamine B in excitation channel 530 with D = 426.4 µm²/s at 298 K and a structural parameter of S = 4, and Atto-655NHS ester in excitation channel 640 with D = 403.6 µm²/s at 298 K and a structural parameter of S = 4). All measurements were performed 20 µm away from the coverslip.

Zelmer C., Zweifel L.P., Kapinos L.E., Craciun I., Güven Z.P., Palivan C.G, & Lim R.Y. (2020). Organelle-specific targeting of polymersomes into the cell nucleus. Proceedings of the National Academy of Sciences of the United States of America, 117(6), 2770-2778.

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2D-STED Imaging of Bacterial Nucleoids

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STED images were recorder with a custom-built STED setup, previously described (53 ). Excitation of the dyes was done with pulsed diode lasers; at 561 nm (PDL561, Abberior Instruments), 640 nm (LDH-D-C-640, PicoQuant) and 510 nm (LDF-D-C-510, PicoQuant). A laser at 775 nm (KATANA 08 HP, OneFive) was used as the depletion beam, which was split into two orthogonally polarized beams that were separately shaped to a donut and a top-hat respectively in the focal plane using a spatial light modulator (LCOS-SLM X10468–02, Hamamatsu Photonics). The laser beams were focused onto the sample using a HC PL APO 100×/1.40 Oil STED White objective (15506378, Leica Microsystems), through which the fluorescence signal was also collected. The images were recorder with a 561nm excitation laser power of 8–20 μW, a 640 nm excitation laser power of 4–10 μW and a 775 nm depletion laser power of 128 mW, measured at the first conjugate back focal plane of the objective. Two-color STED imaging was done in a line-by-line scanning modality. The pixel size was set between 20 and 30 nm with a pixel dwell time of 50 μs. Volumetric 2D-STED imaging of nucleoids was recorded with a voxel size for xyz volumes was set to 25 × 25 × 200 nm³. The pixel dwell time was set at either 30 or 50 μs.

Hirabayashi Y., Lewis TL J.r., Du Y., Virga D.M., Decker A.M., Coceano G., Alvelid J., Paul M.A., Hamilton S., Kneis P., Takahashi Y., Gaublomme J.T., Testa I, & Polleux F. (2024). Most axonal mitochondria in cortical pyramidal neurons lack mitochondrial DNA and consume ATP. bioRxiv.

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