All QDs were purified by precipitation performed by the addition of 2-propanol followed by centrifugation (10303 g for 5 min). The pellet was redissolved in H2O. For csQD-DNA, they were purified by centrifugation through vivaspin centrifugal filter with a 300 kDa molecular weight cut off (3 times centrifugation at 10303 g for 2 min). The photoluminescence decay curves were recorded using the time-correlated single-photon counting (TCSPC) technique on Fluorolog-3 spectrometer (Horiba Jobin Yvon) and the excitation source was a pulsed light emitting diodes with an excitation wavelength of 455 nm (NanoLED-05A, >1.3 ns pulse, Horiba). The curves were fitted with DAS6 analysis software provided by Horiba Scientific. For measurements of photoluminescence quantum yields (PLQY) a G8 integrated sphere module (GMP SA, Switzerland) was used in the Fluorolog-3 spectrometer (Horiba Jobin Yvon).
Fluorolog 3 spectrometer
Manufactured by Horiba
Sourced in United States, Japan, Germany
The Fluorolog-3 is a high-performance spectrofluorometer designed for fluorescence analysis. It features a Xenon lamp source, monochromators for excitation and emission, and a photomultiplier tube detector. The Fluorolog-3 is capable of measuring excitation and emission spectra, as well as time-resolved fluorescence measurements.
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Lab products found in correlation
Cary 6000i spectrophotometer, by Agilent Technologies (2 mentions)
Ti e pfs fluorescence microscopy, by Olympus (2 mentions)
Ix70 inverted microscope frame, by Olympus (2 mentions)
D8 advance, by Bruker (2 mentions)
Quartz cuvette, by Hellma (2 mentions)
Gcr 150 30 nd yag laser, by Spectra-Physics (2 mentions)
Das6 analysis software, by Horiba (1 mentions)
V 670 spectrometer, by Jasco (1 mentions)
Av 400, by Bruker (1 mentions)
Drx 500, by Bruker (1 mentions)
Chi660e, by Chenhua (1 mentions)
Uv 3600 spectrophotometer, by Shimadzu (1 mentions)
Fluoromax 4 spectrometer, by Horiba (1 mentions)
Quanta φ, by Horiba (1 mentions)
Fls980 photoluminescence spectrometer, by Edinburgh Instruments (1 mentions)
Uv 3600 uv vis nir spectrophotometer, by Shimadzu (1 mentions)
Delta flex system, by Horiba (1 mentions)
Uh4150, by Hitachi (1 mentions)
Tecnai f20, by Thermo Fisher Scientific (1 mentions)
Jsm 6700f, by JEOL (1 mentions)
36 protocols using fluorolog 3 spectrometer
1
Purification and Characterization of Quantum Dots
2
Optical Characterization of Ir(III) Complexes
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The ultraviolet-visible (UV-vis) absorption spectra were taken using the JASCO V670
spectrometer. The photoluminescence (PL) spectra were recorded using the Horiba Fluorolog 3
spectrometer with a Xe lamp as the excitation source. The phosphorescence spectra of Ir complexes were also recorded using Horiba Fluorolog 3 spectrometer equipped with liquid nitrogen dewar assembly. The exciton lifetime of Ir(III) complexes were measured using a time-
spectrometer. The photoluminescence (PL) spectra were recorded using the Horiba Fluorolog 3
spectrometer with a Xe lamp as the excitation source. The phosphorescence spectra of Ir complexes were also recorded using Horiba Fluorolog 3 spectrometer equipped with liquid nitrogen dewar assembly. The exciton lifetime of Ir(III) complexes were measured using a time-
3
Absolute Fluorescence Quantum Yield Measurement
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The absolute fluorescence quantum yield was measured on a Horiba Fluorolog-3 spectrometer (Edison, NJ, USA) equipped with an integrating sphere. A xenon lamp coupled to a double monochromator was used as the excitation light source. The sample (1 cm quartz cuvette cell with solution in THF) or blank (pure THF) was directly illuminated in the center of the integrating sphere. The optical density of all investigated sample solutions in corresponding solvents did not exceed 0.1 at the luminescence excitation wavelength. Under the same conditions (e.g., excitation wavelength, spectral resolution, and temperature), the luminescence spectrum of the sample Ec, the luminescence spectrum of the blank Ea, the Rayleigh scattering spectrum of the sample Lc, and the Rayleigh scattering spectrum of the solvent La were measured. The absolute fluorescence quantum yield was determined according to the formula:
4
Characterization of Organic Compounds
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Elemental analysis was performed with a EuroEA3000 analyzer using standard technique. The IR spectra were recorded in KBr on a Bruker Vector-22 spectrometer.1H and 13C NMR spectra were recorded on Bruker AV-400 (400.13 and 100.61 MHz) and Bruker DRX-500 (500.13 and 125.76 MHz) spectrometers using the residual signals of the solvent (CDCl3) at 7.24 ppm for 1H and 76.9 ppm for 13C with respect to TMS as the internal standard. Corrected photoluminescence spectra were recorded on a Fluorolog 3 spectrometer (Horiba Jobin Yvon).
5
Protein-ligand Binding Kinetics by Fluorescence
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Fluorescence quenching and fluorescence anisotropy measurements were carried out in triplicates at 25°C on a Fluorolog 3 spectrometer (Horiba Jobin Yvon) equipped with polarizers. For fluorescence quenching with SAM, SAH and 5′-methylthioadenosine experiments the tryptophan fluorescence of SsTsr3 (200 nM in 25 mM Tris–HCl pH 7.8, 250 mM NaCl, 2 mM β-mercaptoethanol) was excited at 295 nm and emission spectra were recorded from 250 to 450 nm for each titration step. The fluorescence intensity at 351 nm for each titration step was normalized with regard to the fluorescence of the free protein and was used for deriving binding curves. KD's were derived by nonlinear regression with Origin 8.0 (Origin Labs) using Equation (1 ):
(F is the normalized fluorescence intensity, a is the change in fluorescence intensity, c is the ligand concentration and KD is the dissociation constant).
5′-Fluoresceine labeled RNAs for fluorescence anisotropy measurements were obtained commercially (Dharmacon), deprotected according to the manufacturer's protocol and the RNA concentration adjusted to 50 nM in 25 mM Tris–HCl pH 7.8, 250 mM NaCl. Fluoresceine fluorescence was excited at 492 nm and emission was recorded at 516 nm. The data were fitted to Equation (1 ) (F is the normalized fluorescence anisotropy, a is the change in fluorescence anisotropy).
(F is the normalized fluorescence intensity, a is the change in fluorescence intensity, c is the ligand concentration and KD is the dissociation constant).
5′-Fluoresceine labeled RNAs for fluorescence anisotropy measurements were obtained commercially (Dharmacon), deprotected according to the manufacturer's protocol and the RNA concentration adjusted to 50 nM in 25 mM Tris–HCl pH 7.8, 250 mM NaCl. Fluoresceine fluorescence was excited at 492 nm and emission was recorded at 516 nm. The data were fitted to Equation (
6
Absorption and Emission Spectroscopy
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The absorption spectra were measured on an Agilent 8453 UV–VS Spectrometer. Steady-state emission experiments and lifetime measurements were performed on a Horiba Jobin Yvon FluoroLog-3 spectrometer. Low temperature (77 K) emission spectra and lifetimes were measured in 2-MeTHF cooled with liquid nitrogen.
7
Fluorescence Characterization of Adhesive Fibers
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Fluorescence images were recorded on a Nikon Ti-E PFS fluorescence microscopy coupled with an Olympus IX70 inverted microscope frame (Olympus UK, Essex, UK). Ultraviolet-Visible (UV-Vis) absorption spectra were recorded with a CARY-6000i spectrophotometer. Fluorescence emission and excitation spectra were measured using a FluoroLog 3 spectrometer manufactured by HORIBA Jobin Yvon. The fluorescence quantum yields (Q (%)) of adhesive fibers were assessed with the Williams comparative method26 using a standard blue fluorophore (Coumarin 102) with a known quantum yield of Q = 76.4% in ethanol. Detailed information about determination of fluorescence quantum yields are described in Supplementary Section 14 and Supplementary Figure 17 .
8
Fluorescence Characterization of Adhesive Fibers
9
Liposome Membrane Fluidity Analysis
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The membrane fluidity of the different liposomes was assessed by steady-state fluorescence anisotropy of diphenyl-hexatriene (DPH). For that, 600 μL of the previously prepared liposome suspension was transferred to a quartz cuvette containing 2400 μL PBS. The liposomes were labelled with DPH (1 μL of 5 mM stock solution in tetrahydrofuran) in a dark at 25 °C for 5 min.
Steady-state anisotropy of DPH was measured in a Fluorolog-3 spectrometer (Horiba Jobin Yvon) using T-format fluorescence polarizers. During the measurements, samples were stirred and equilibrated in a temperature-controlled chamber using a thermoelectric Peltier junction. The excitation and emission wavelengths were 360 nm (5 nm bandwidth) and 431 nm (5 nm bandwidth), respectively. Steady-state fluorescence anisotropy (r) was calculated as follows:
where I is the fluorescence intensity, and the first and second subscripts refer to the setting of the excitation and emission polarizers, respectively. is a correction factor for the monochromator’s transmission efficiency for vertically and horizontally polarized light.
Steady-state anisotropy of DPH was measured in a Fluorolog-3 spectrometer (Horiba Jobin Yvon) using T-format fluorescence polarizers. During the measurements, samples were stirred and equilibrated in a temperature-controlled chamber using a thermoelectric Peltier junction. The excitation and emission wavelengths were 360 nm (5 nm bandwidth) and 431 nm (5 nm bandwidth), respectively. Steady-state fluorescence anisotropy (r) was calculated as follows:
where I is the fluorescence intensity, and the first and second subscripts refer to the setting of the excitation and emission polarizers, respectively. is a correction factor for the monochromator’s transmission efficiency for vertically and horizontally polarized light.
10
Characterization of Photocatalytic Nanomaterials
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The XRD patterns were obtained using an X-ray diffraction diffractometer (Bruker D8-Advance, Germany) with Cu-Kα radiation. TEM images of the samples were recorded through the transmission electron microscopy (JEM-2100, Japan). Surface chemical states of the photocatalysts were measured by X-ray photo-electron spectroscopy (XPS, AXISULTRA) with monochromatic Al Ka X-rays (1486.6 eV). The photoluminescence (PL) spectra were measured on a JY HORIBA FluoroLog-3 spectrometer, and the excited wavelength of 460 nm was chosen. The curves of photocurrent response were carried out in an electrochemical workstation (CHI660E, Chenhua, China) using a conventional standard three-electrode cell under visible light irradiation (λ > 420 nm). 0.1 mol/L Na2SO4 solution was used as electrolyte.
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