Fluorocube

Manufactured by Horiba

Sourced in Japan

The Fluorocube is a compact fluorescence spectrometer designed for measurement of fluorescent samples. It provides accurate and reliable fluorescence intensity data across a wide wavelength range. The core function of the Fluorocube is to excite and detect fluorescence emissions from samples.

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Lab products found in correlation

Nanoled 293, by Horiba (2 mentions) Fluoromax 4 spectrofluorometer, by Horiba (1 mentions) C9920 02 system, by Hamamatsu Photonics (1 mentions) Fp 8500, by Jasco (1 mentions) C10027, by Hamamatsu Photonics (1 mentions) C9920, by Hamamatsu Photonics (1 mentions) V 670, by Jasco (1 mentions) Vg escalab 220i, by Thermo Fisher Scientific (1 mentions) Jem 2100f, by JEOL (1 mentions) Labram aramis, by Horiba (1 mentions) Dimethyl sulfoxide (dmso), by Fujifilm (1 mentions) Avance 3 500 nmr instrument, by Bruker (1 mentions) Du800 spectrophotometer, by Beckman Coulter (1 mentions) Ls50b fluorescence spectrometer, by PerkinElmer (1 mentions) Dmso d6, by Merck Group (1 mentions) Ludox, by Merck Group (1 mentions)

Close spelling variants of this product

Fluorocube-01-NL (5 citations) FluoroCube fluorimeter (3 citations) Fluorocube-01-NL lifetime system (2 citations) FluoroCube time-correlated single photon counting system (2 citations)

13 protocols using fluorocube

Trp Fluorescence Anisotropy Decay Analysis

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The fluorescence properties of Trp in proteins were probed by exciting at 295 nm and monitoring the emission for 310–450 nm (λ_em = 338 nm) at 25 °C using Jobin Yvon Fluoromax‐4 spectrofluorometer equipped with a PMT detector. The slit width (2 nm), step size (0.1 nm), and integration time (0.05 s) were maintained for all experiments.
Fluorescence anisotropy decay measurements were performed using TCSPC (Fluorocube, Horiba Jobin Yvon, Kyoto, Japan). For decay measurements, the emission polarizer was set to 0° with respect to excitation polarizer for parallel measurements and at 90° for perpendicular measurements. A 293 nm laser diode was used as an excitation source, and the emission monochromator for tryptophan was fixed at 342 nm, at a slit width of 8 nm. The instrument response function was measured using 2% LUDOX (Sigma‐Aldrich).
The anisotropy decay was calculated using Equation (2) and was fitted to biexponential decay fit to determine the values of rotational correlation times using Equation (3). The values are tabulated in Table S6.

r (t) = \frac{I_{⊥} (t) - I_{‖} (t)}{I_{⊥} (t) + 2 I_{‖} (t)}

r_{t} = r_{0} [A_{1} e^{\frac{- t}{ϕ 1}} + A_{2} e^{\frac{- t}{ϕ 2}}]

where I_⊥ is the vertical emission and I_‖ is the horizontal emission.

Singaraju G.S., Sagar A., Kumar A., Samuel J.S., Hazra J.P., Sannigrahi M.K., Yennamalli R.M., A.s.h.i.s.h, & Rakshit S. (2019). Structural basis of the strong cell‐cell junction formed by cadherin‐23. The Febs Journal, 287(11), 2328-2347.

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Absorption, Emission, and Lifetime Spectroscopy

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UV/Vis absorption spectra were recorded on a PerkinElmer (Lambda 750) UV-VIS-NIR spectrophotometer. Fluorescence and phosphorescence emission spectra were recorded on a PerkinElmer (LS-55) spectrofluorophotometer. Fluorescence lifetimes were measured on a HORIBA Scientific time-correlated single-photon counting system (FluoroCube) with the laser light (DeltaDiode, laser diode head, 404 nm, pulse width: 100 ps) as the excitation source. Phosphorescence lifetimes were measured on a JASCO FP-8500. The absolute fluorescence quantum yields were determined by a Hamamatsu Photonics C9920-02 system equipped with an integrating sphere and a red-sensitive multichannel photodetector (PMA-12): excitation wavelength = 300 nm.

Aoki T., Sakai H., Ohkubo K., Sakanoue T., Takenobu T., Fukuzumi S, & Hasobe T. (2014). Ultrafast photoinduced electron transfer in face-to-face charge-transfer π-complexes of planar porphyrins and hexaazatriphenylene derivatives †Electronic supplementary information (ESI) available: 1H, 13C NMR and MALDI-TOF mass spectra, cyclic voltammograms, fluorescence spectra, fluorescence titration spectra, 1H NMR titration, femtosecond laser-induced transient absorption spectral measurement data, and DFT data. See DOI: 10.1039/c4sc02787f Click here for additional data file.. Chemical Science, 6(2), 1498-1509.

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Time-Resolved Fluorescence Spectroscopy

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Fluorescence decay lifetimes were measured using a time-correlated single photon counting instrument (Fluorocube, Horiba JobinYvon, Edison, NJ) with a 457 nm pulsed LED (NanoLED-293, Horiba JobinYvon, Edison NJ) light source having a 1 ns pulse width and 1 MHz repetition rate. Decays were recorded at the emission maximum of each PDI with a 32 nm bandwidth.

Farooqi M.J., Penick M.A., Burch J., Negrete G.R, & Brancaleon L. (2015). Characterization of novel perylene diimides containing aromatic amino acid side chains. Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy, 153, 124-131.

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Fluorescence Lifetime Analysis of QBA-DNA Interactions

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QBAs in 3 × 10^-6M concentration were prepared in 0.1M borate buffer, pH 9.45 (alkanolamine form) or in 0.1M acetate buffer, pH 3.95 (iminium form). For measurement of lifetimes of QBA-DNA complexes, ctDNA was added (DNA base pair:drug ratio 1.6:1). Measurements were performed on a Fluorocube (Horiba Jobin Yvon, France) with a 329 nm excitation LED diode (pulse width 1.2 ns) and emission monochromator set to 440 nm (alkanolamine form) or 610 nm (iminium form, 570 nm for chelerythrine). Instrument response function was obtained by measuring a solution of colloidal silica. When necessary, intensity was lowered by neutral filters. For each QBA three measurements were performed and lifetimes were globally fitted using DecayFit 1.3. Quality of a fit was considered by inspecting residuals and reduced χ² value around 1. SE are reported in Table 1.

Rájecký M., Šebrlová K., Mravec F, & Táborský P. (2015). Influence of Solvent Polarity and DNA-Binding on Spectral Properties of Quaternary Benzo[c]phenanthridine Alkaloids. PLoS ONE, 10(6), e0129925.

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Fluorescence Lifetime Measurement of HSA

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Fluorescence decay lifetimes were measured using a time-correlated single photon counting (TCSPC) instrument (Fluorocube, Horiba Scientific, Edison, NJ) with a 294 nm pulsed LED (NanoLED-293, Horiba JobinYvon, Edison NJ) source of ~1 ns pulse width and 1 MHz repetition rate. Decays were recorded at the emission maximum of HSA with a 12 nm bandwidth.

Rozinek S.C., Thomas R.J, & Brancaleon L. (2016). Biophysical characterization of the interaction of human albumin with an anionic porphyrin. Biochemistry and Biophysics Reports, 7, 295-302.

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Spectroscopic Characterization of Fluorescent Proteins

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The absorption
spectra were recorded with a spectrophotometer (V-670, Jasco). The
fluorescence spectra were recorded using a photonic multi-channel
analyzer (C10027, Hamamatsu Photonics). The fluorescence decay curves
of EGFP emissions were measured by excitation at 485 nm using a time-correlated
single-photon counting kit (Horiba Fluoro Cube). The fluorescence
autocorrelation curves were measured on a compact FCS system (C9413-01MOD,
Hamamatsu Photonics, Japan) at an excitation of 473 nm using a laser-diode-pumped
solid-state laser. The size of the pinhole was 25 μm, and the
spectral range of the detection wavelengths was 500–900 nm.
For the determination of the concentration of GSH–QDs, the
number of QD particles in a 10 μL solution was measured by using
FCS, and the QD concentration was estimated by using a 20 nM solution
of rhodamine 6G as a reference. Fluorescence quantum yields were measured
using an absolute quantum yield measurement system (C9920, Hamamatsu
Photonics). Excitation wavelengths were set to be 475 nm for annexin
V–EGFP and 488 nm for GSH–QDs.

Tsuboi S, & Jin T. (2022). In Vitro and In Vivo Fluorescence Imaging of Antibody–Drug Conjugate-Induced Tumor Apoptosis Using Annexin V–EGFP Conjugated Quantum Dots. ACS Omega, 7(2), 2105-2113.

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Fluorescence Lifetime Spectroscopy of Tubulin

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Fluorescence lifetime was recorded using a time-correlated single-photon-counting (TCSPC) instrument (Fluorocube, Horiba Scientific, Edison, NJ). The details of the method can be found elsewhere ^{[21 , 38 (link)]} and are only summarized in the following: tubulin fluorescence decay was recorded upon excitation with a pulsed LED at 294 nm (NanoLED-295, 1.2 ns pulsewidth, Horiba Scientific), operated at a repetition rate of 1.0 MHz. The instrument response function (prompt) was recorded after each decay curve using a suspension of glycogen (1 mg/mL) in deionized water. All signals were recorded by accumulating 10⁴ peak counts at a counting rate below the suggested maximum (2 × 10⁴ cts/s) corresponding to 2% of the repetition rate of the source. The decays shown in Figure 4 and 9 are plots of the logarithm of the number single photons counted as a function of the time delay from the trigger of the pulse of the Nano-LED source.

Sagarra A.V., McMicken B., Nonell S, & Brancaleon L. (2016). Effects of Visible Irradiation of Protoporphyrin IX on the Self Assembly of Tubulin Heterodimers. Chemphyschem : a European journal of chemical physics and physical chemistry, 17(20), 3269-3282.

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Characterization of Graphene Quantum Dots

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HR-TEM images were taken using a 2100F field emission gun TEM (JEM 2100F, JEOL, USA, 200 kV) for on-GQDs and off-GOQDs samples. XPS spectra were recorded for both samples using VG ESCALAB 220i (Thermo scientific, USA). XPS survey and high-resolution scans were performed with the pass energies of 100 eV and 20 eV, respectively. X-ray beam size was approximately 100 μm. On-GQDs and off-GOQDs samples for XPS measurement were prepared via a spin coating technique. Silicon (Si) substrate was used for spin coating. The rotation speed was adjusted to 3000 rpm. The samples were dried at room temperature for 2 h before the measurement. Room temperature PL spectra of on-GQDs and off-GOQDs were collected using a photoluminescence spectrophotometer (Horiba, Fluoromax-plus with 150 W xonon arc lamp) in the wavelength range from 300 nm to 700 nm. Raman spectra were obtained for both samples using Raman spectrometer (LabRam Aramis, Horiba jobin Yvon, 514 nm Ar-ion laser used with a power of 1 mW). A time-correlated single photon counting (TCSPC) spectrometer (Horiba Jobin Yvon, FluoroCube) was used to measure nanosecond lifetime.

Kang S., Jeong Y.K., Jung K.H., Son Y., Choi S.C., An G.S., Han H, & Kim K.M. (2019). Simple preparation of graphene quantum dots with controllable surface states from graphite. RSC Advances, 9(66), 38447-38453.

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Photophysical Characterization of Organic B

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UV–vis measurements were performed in transmission mode using an AvaLight-DHS-Bal (Balanced Deuterium-Halogen light source (200−2000 nm)). Extinction spectra of B dissolved in toluene were recorded on a jasco UV/Vis V-670 spectrophotometer. Photoluminescence spectra including the derived quantum yields were measured under current nitrogen flow using a PTI QuantaMaster 40 spectrofluorometer equipped with an integrated sphere (Labsphere) and a Hamamatsu R928P photomultiplier. The samples were excited at wavelengths of 355, 375, and 410 nm with a Xenon lamp using a monochromator. The TRPL spectra were recorded under current nitrogen flow using a time-correlated single photon counting (TCSPC) system (Horiba Fluorocube). TRPL samples were excited at 375 nm with a pulsed laser diode (Horiba NanoLED Model N-375L).

Heimel P., Mondal A., May F., Kowalsky W., Lennartz C., Andrienko D, & Lovrincic R. (2018). Unicolored phosphor-sensitized fluorescence for efficient and stable blue OLEDs. Nature Communications, 9, 4990.

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Synthesis and Characterization of 1-Pyrenebutyric Acid N-Hydroxysuccinimide Ester

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Unless noted otherwise, all reagents were obtained from commercial sources and used without further purification. The 1-pyrenebutyric acid N-hydroxysuccinimide ester was obtained from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). Dimethyl sulfoxide was obtained from Fujifilm Wako Pure Chemical Corporation (Osaka, Japan). DMSO-d₆, was obtained from Sigma-Aldrich Inc. (St. Louis, MO, USA). UV absorption spectroscopy was performed using a DU-800 spectrophotometer (Beckman-Coulter, Brea, CA, USA). Fluorescence spectra were measured using a LS50B fluorescence spectrometer (Perkin Elmer, Waltham, MA, USA). Time-resolved fluorescence measurement was performed using a single photon counter FluoroCube (Horiba, Kyoto, Japan). ¹H NMR spectra were recorded using an Avance III-500 NMR instrument (Bruker, Billerica, MA) at the Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST). HRMS was performed by the Open Research Facilities Station, TIA Central Office, AIST.

Naya M, & Sato C. (2020). Pyrene Excimer-Based Fluorescent Labeling of Cysteines Brought into Close Proximity by Protein Dynamics: ASEM-Induced Thiol-Ene Click Reaction for High Spatial Resolution CLEM. International Journal of Molecular Sciences, 21(20), 7550.

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