Rf 6000 fluorescence spectrophotometer

The db/db mice received ACLT surgery were randomly divided into three groups (DiR‐Vt, DiR‐VPs, and DiR‐MVPs). Then, the mice were intravenously injected with different formulations (DiR equivalent to 0.1% of Vt, the concentration of DiR was ≈0.05 µm). The fluorescence distribution of the right knees was measured at 0, 1, 2, 3, 4, 6, and 9 days after injection by RF‐6000 fluorescence spectrophotometer (Shimadzu, Japan). For the bio‐distribution of MVPs, organ distribution was also measured accordingly.

All analytical-grade chemicals used in this study were purchased from Aladdin Chemistry Co., Ltd. (Shanghai, China) and used directly without purification. HRP (≥300 U·mg⁻¹) was purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China). H₂O was purified using a Milli-Q water purification system (Millipore, St. Louis, MO, USA). The UV–Vis and fluorescence spectra of the nanomaterials were collected using a UV-2700 spectrophotometer (Shimadzu Corp., Kyoto, Japan) and an RF-6000 fluorescence spectrophotometer (Shimadzu Corp.), respectively. FTIR spectra were acquired using a VERTEX 80VFT-IR spectrometer (Bruker Daltonik GmbH, Bremen, Germany). The TEM images were obtained using a JEM-2100 microscope (JEOL Ltd., Tokyo, Japan) at 200 kV. The particle sizes were determined using the Dynamic Light Scattering (DLS) method with a Zetasizer Nano ZS 90 (Malvern Panalytical, Malvern, UK).

The formation of the PilB_1-190-PilZ_{W69_5OHW} and FimX_EAL-PilZ_{W69_5OHW} complexes were monitored by changes in the 5-hydroxytryptophan fluorescence emission spectra of PilZ_{W69_5OHW} upon addition of different amounts PilB_1-190 or FimX_EAL. The assay was carried out in buffer containing 50 mM Tris-HCl pH 8,0, 50 mM NaCl, 1 mM MgCl₂, 1 mM β-mercaptoethanol and the initial concentrations of PilZ_{W69_5OHW} was 1 μM. The samples were equilibrated for 2 min before each measurement. Titration experiments were performed using a RF-6000 fluorescence spectrophotometer (SHIMADZU). The excitation wavelength was 310 nm (bandwidth: 5 nm), and the emission spectra were recorded between 325–445 nm (bandwidth: 5 nm). Dissociation constants were calculated assuming a simple 1:1 bonding model, as previously described [82 (link)] using the SigmaPlot 11 software (Systat Software Inc.).

RF‐6000 fluorescence spectrophotometer (Shimadzu, Japan) was employed to investigate the intrinsic fluorescence emission profile of rhFTL in PBS buffer. The protein sample was diluted to about 0.1 mg/mL and the path length of the cuvette was 1.0 cm. The excitation wavelength was constant at 280 nm, and fluorescence emission signals were recorded from 260 to 450 nm.

TD-3500 X-ray diffractometer (XRD) for determination of crystal structure of samples. The measurement conditions were using Cu-Kα as a source (λ = 0.154 nm), a scanning speed of 5 °/min, voltage of 40 kV, and current of 30 mA. The morphology and microstructure of the samples were obtained using field emission scanning electron microscopy (FE-SEM) (JSM-7100F) and transmission electron microscopy (TEM) (JEM-2010) at 200 kV. The composition and content of elements in the sample were determined by energy dispersive X-ray energy spectrometer (EDS) on scanning electron microscope. The UV–Vis diffuse reflectance spectrum was obtained on a Hitachi U-3900 U–Vis spectrophotometer. The instrument uses barium sulfate as reference material. Shimadzu RF-6000 fluorescence spectrophotometer obtained photoluminescence (PL) spectra of samples. The Zeta potential measurement was measured as follows: 5 mg of g-C₃N₄, Zn₃In₂S₆, and Zn₃In₂S₆/g-C₃N₄ nanocomposite was added to 10 mL of distilled water and then sonicated for 15 min (40 kHz, 600 W bath type sonicator), respectively. Further, 1 mL of suspension was injected into the cell for the determination of zeta potential (Nano ZS90 Malvern, UK).

Reagents related to the synthesis of Cu[GluC] were commercial reagents without further purification and were provided in the Supplementary Material. The water used in this work was ultrapure water. Sodium, potassium, and copper salts were purchased from Adamas Chemical Reagent Co. Stock solutions of these salts were prepared with ultrapure water, which was also used throughout the study. The synthetic compounds were characterized with a nuclear magnetic resonance (NMR) spectrometer (Bruker, Karlsruhe, Germany) where the residual signals from DMSO-d₆ (¹H: δ 2.50 ppm) or Chloroform-d (¹H: δ 7.26 ppm) were used as internal standards and High-resolution ion mobility liquid chromatography–mass spectrometry (HRLCMS) (LC-30A + TripleTOF5600+, AB SCIEX, USA). The UV-vis spectra were measured with a 1750 UV-visible spectrophotometer (Shimadzu, Japan), and the fluorescence spectra were measured with an RF-6000 fluorescence spectrophotometer (Shimadzu, Kyoto, Japan).