The fusion of R18-TOSV with host cell membranes was performed as previously described [34 (link)]. Briefly, cells were detached from the culture surface using 0.5 mM EDTA, and binding of R18-TOSV at MOI 10 to cells in suspension was synchronized on ice in a phenol-free medium at pH ∼7.4 for 90 min. To determine the kinetics of virus penetration, virus-bound cells were rapidly warmed in an FP-8500 fluorometer (Jasco) to 37°C, and the emission of fluorescence was measured over 90 min. For virus fusion with cell membranes, virus-bound cells were rapidly warmed inside the fluorometer to 37°C, and the fusion was triggered with buffers of varying pHs as indicated. The fluorescence was measured for 600 sec. Where indicated, virions were pre-exposed to buffers at pH ∼7.4 or 6.0 as described in the Virus binding and internalization section.
Fp 8500 fluorometer
Manufactured by Jasco
The FP-8500 is a fluorometer designed for the measurement of fluorescence intensity. It is capable of performing excitation and emission scans to determine the optimal wavelengths for fluorescence analysis. The FP-8500 is a versatile instrument that can be used for a variety of applications requiring the quantification of fluorescent signals.
Automatically generated - may contain errors
Lab products found in correlation
Spectra manager 2 software, by Jasco (3 mentions)
S 3100 spectrophotometer, by Scinco (2 mentions)
Fp 8500 fluorescence spectrometer, by Jasco (1 mentions)
Quantaurus qy spectrometer, by Hamamatsu Photonics (1 mentions)
V 500 absorption spectrometer, by Jasco (1 mentions)
F 7000, by Hitachi (1 mentions)
7 protocols using fp 8500 fluorometer
1
Kinetics of TOSV Membrane Fusion
2
Spectroscopic Analysis of Photoluminescence
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UV–vis absorption spectra
were recorded using a JASCO V-500 absorption spectrometer (JASCO,
Japan). Steady-state PL spectra were obtained using a Hitachi F-7000
(Hitachi, Japan) or a JASCO FP-8500 fluorescence spectrometer (JASCO,
Japan). PL quantum yields (PLQYs) were estimated using a calibrated
integrating sphere system attached to a JASCO FP-8500 fluorometer.
PLQYs during the heating/cooling processes were measured by using
a Quantaurus-QY spectrometer (Hamamatsu Photonics K.K., Japan).
were recorded using a JASCO V-500 absorption spectrometer (JASCO,
Japan). Steady-state PL spectra were obtained using a Hitachi F-7000
(Hitachi, Japan) or a JASCO FP-8500 fluorescence spectrometer (JASCO,
Japan). PL quantum yields (PLQYs) were estimated using a calibrated
integrating sphere system attached to a JASCO FP-8500 fluorometer.
PLQYs during the heating/cooling processes were measured by using
a Quantaurus-QY spectrometer (Hamamatsu Photonics K.K., Japan).
3
Absolute Quantum Yield Measurement
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2 mm × 10 mm QS-grade quartz cuvettes (Jasco Parts Center 6808-H250A) were used. Absolute quantum yields were recorded on a Jasco FP-8500 fluorometer equipped with a 100-mm integrating sphere setup (ILF-835), and calculated using Jasco Spectra Manager II Software.
4
Quartz Cuvette Spectroscopic Analysis
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10 mm × 10 mm QS-grade quartz cuvettes (Hellma Analytics 111-QS) were used. Hexane was used as a solvent. Light absorption spectra were recorded on a Scinco S-3100 spectrophotometer. Photoluminescence spectra were recorded on a Jasco FP-8500 fluorometer.
5
Fabrication and Optical Characterization of C-dot Films
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C-dots (250 mg) were dissolved in toluene (5 ml). The solution (5 ml) was mixed with the 10% poly(methyl methacrylate) solution in anisole (5 ml). The mixture was vortexed for 10 min and air bubbles were removed. The mixture (0.5 ml) was then dropped onto 18 mm × 18 mm cover glasses and kept undisturbed for 24 h on a flat table. Once the solvent was evaporated completely, the resulting films were peeled off from the cover glasses and placed over 280–320 nm InGaN light-emitting diodes by using adhesive tapes. Emission spectra were recorded on a Minolta CS2000 spectroradiometer with a Keithley 236 sourcemeter. Internal quantum yields were recorded on a Jasco FP-8500 fluorometer equipped with a 100 mm integrating sphere setup (ILF-835) and calculated by using Jasco Spectra Manager II Software.
6
Absorption and Photoluminescence Spectroscopy
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Light absorption spectra were recorded on a Scinco S-3100 spectrophotometer. Photoluminescence excitation and emission spectra were recorded on a Jasco FP-8500 fluorometer.
7
Quantum Yield Measurement Protocol
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2 mm × 10 mm QS-grade quartz cuvettes (Jasco Parts Center 6808-H250A) were used. Absolute quantum yields were recorded on a Jasco FP-8500 fluorometer equipped with a 100 mm integrating sphere setup (ILF-835) and calculated by using Jasco Spectra Manager II Software.
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