Fs5 spectrofluorometer

Protein endogenous fluorescence spectra of OVM, OVM-COS, and OVM-COS-LUT were measured using a fluorescence spectrophotometer (Spectrofluorometer FS5, Edinburgh Instruments, Livingston, UK). The sample protein concentration was diluted to 0.5 mg/mL; the excitation wavelength was 274 nm. The wavelength scan range was 280–500 nm; the excitation and emission slit widths were set to 2 nm.

High‐angle annular dark‐field imaging scanning transmission electron microscope (HAADF‐STEM) images were acquired on an FEI Titan Cubed Themis G201 double spherical aberration‐corrected transmission electron microscope with an acceleration voltage of 300 kV. Absorbance spectra were measured using a GBC Cintra2020 spectrometer with tunable wavelengths. Photoluminescence (PL) spectra were obtained on Spectrofluorometer FS5 (Edinburgh instruments). Scanning electron microscope (SEM) images were obtained using a HITACHI SU8010 scanning electron microscope. Electrical characteristics of the transistors were measured using the combination of a Keithley 4200SCS parameter analyzer and a probe station. CHI continuous lasers with wavelengths of 405 or 637 nm were used for the relevant experiments. The electrical and optical measurements conducted on the devices were carried out in a glovebox filled with nitrogen.

Fluorimetry was done using Edinburgh Instruments Spectrofluorometer FS5 and the data collected with Fluoracle software. A literature method was followed^{45 (link)}, using 1,6-diphenyl-1,3,5-hexatriene (DPH) as a fluorescent probe with excitation wavelength of 358 nm and emission wavelength of 430 nm. The CMC for alkenes 3a–d was extracted from a plot of emission vs. concentration (Supplementary Figs. 39–42). Experiments were performed at 60 °C to stay within optimal instrument parameters and the trends observed at 60 °C are expected to hold under reaction conditions at 80 °C.

Powder X‐ray diffraction (XRD) of InP/ZnS core/shell QDs was carried out using a Rigaku Miniflex XRD λ_{Cu Kα} = 1.54 Å with a 5° min⁻¹ scan rate. XRD measurements were carried out at room temperature. XRD samples were prepared by drop‐casting QDs dissolved in hexane onto 1 cm × 1 cm silicon wafers and heated to 100 °C and kept at this temperature for 1 h to dry liquid QDs on the silicon wafers. DF‐STEM images were collected using Hitachi HF5000 TEM/STEM operated at 200 kV. For TEM measurement, samples were deposited as 10 µL of a 1 mm QD in a hexane solution on a copper support grid.
An Edinburgh Instruments spectrofluorometer FS5 with a 150 W xenon lamp was used to measure absorbance and PL. The excitation wavelength was 375 nm. For absorbance and PL measurements, regular quartz cuvettes were used. PLQY measurements were carried out using an integrating sphere module with an inner diameter of 150 mm. The integrating sphere was inserted into the FS5 device for the measurement of quantum yields.

Coupling Medium: The main component of the isolation coupling pad (icPad) is polyacrylamide with a diameter of 120 mm and a thickness of 8 mm [11 (link)]. There are two sides of icPad, the A-side and B-side; the A-side is attached to the EWSL probe, and the B-side is attached to the patient body (Figure 1).
Contaminant-Simulating Substance: Rhodamine 6G was purchased from Tokyo Chemical Industry Company (Japan). Ethanol was bought from Sigma-Aldrich company (USA).
Equipment: Spectrofluorometer FS5 (Edinburgh Instruments, Livingston, United Kingdom).

The fluorescence anisotropy of FAM-labeled ACR3 oligonucleotides (labeled on the 5′ end with 6-carboxyfluorescein) was measured on two-four independent repetitions with different protein to DNA ratios and one reference solution without protein in buffer A (10 mM Tris–HCl pH 8, 50 mM NaCl, 0.5% NP-40, 1 mM DTT and 5% glycerol). The total volume of the working solutions was 100 μl, and the added protein elution buffer amount was kept constant to 50 μl by adding buffer A when necessary. Measurements were performed on a spectrofluorometer FS5 (Edinburgh Instruments) in a temperature-controlled microcuvette at 25°C. Fluorescence emission intensity was recorded at 515 nm, with excitation at 490 nm, and emission and excitation slits set to 2 nm. All titrations were performed using 1 nM of DNA, and after each addition the sample was equilibrated for 6 min. Stoichiometric binding curves were fit to the equation: ΔA = ΔA_T/2D_T{(E_T+D_T+K_d) – [(E_T+D_T+K_d)² – 4E_T+D_T]^1/2}, where ΔA is the change in anisotropy, ΔA_T is the total anisotropy change, E_T is the total protein concentration, D_T is the total DNA concentration, and K_d is the dissociation constant.

The molecular weight (M_w and M_n) and polydispersity (M_w/M_n) of the polymers were estimated in THF using a gel permeation chromatography (GPC, PL-GPC-50, Waters Corporation, Milford, CT, USA) system with a set of monodisperse polystyrene standards covering the molecular weight varying from 10³ to 10⁷ as calibration. FTIR spectra were recorded on a VECTOR 22 (Bruker Corporation, Billerica, MA, USA) spectrometer. ¹H NMR spectra were recorded on AVANCE III 400 (Bruker Corporation, Billerica, MA, USA) spectrometers, and tetramethylsilane (TMS) was used as an internal standard. UV-vis absorption spectra were recorded on a Varian CARY 100 Bio UV-vis (Agilent Technologies Inc, Santa Clara, CA, USA)spectrophotometer. Fluorescence spectra were recorded on a RF-5301PC (Shimadzu, Kyoto, Japan) spectrofluorophotometer. Quantum yield was recorded on the Spectrofluorometer FS5 (Edinburgh Instruments, Livingston, Scotland, UK) spectrofluorophotometer.