Zetasizer ultra instrument

Surface morphology and elemental composition were characterized by a TESCAN MIRA3 XMU SEM equipped with an Oxford Instruments energy dispersive X-ray (EDX) detector. The Brunauer–Emmett–Teller (BET) specific surface area was evaluated from the adsorption data registered through a 3P micro 300 instrument in the relative pressure (p/p₀) range of 0.1 ÷ 0.3. Before BET measurements, samples were degassed at 300 °C overnight^{44 (link)}. The crystalline structure was determined by XRD in parallel beam mode using a Rigaku SmartLab 9 kW diffractometer equipped with a high-brightness Cu K_α rotating anode X-ray tube operated at 45 kV and 150 mA. Surface chemical composition was studied by XPS using a Kratos Analytical Axis Supra instrument with a monochromatic Al K_α (1486.7 eV) excitation source. All spectra were calibrated to the adventitious C 1s peak at 284.8 eV and fitted using CasaXPS software. The magnetic hysteresis loop was measured using a Quantum Design VersaLab cryogen-free VSM at 300 K for an applied magnetic field ranging from −10 to 10 kOe at steps of 10 Oe s⁻¹. Light-absorption spectra were measured using a Jasco V-750 UV–Visible spectrophotometer equipped with an integrating sphere. Zeta potential measurements were performed in water at pH 7 and 3 using a Malvern Panalytical Zetasizer Ultra instrument.

Blood samples were collected from mice fed with a normal diet, mice fed with a high-fat diet, and mice subjected to exercise training after a high-fat diet, and then centrifuged at 10,000g for 30 min and at a speed of 100,000×g for 90 min using a Sorvall MTX150 high-speed centrifuge (46,963, Thermo Fisher Scientific, USA). The samples were resuspended in 1 × PBS, treated with 25 mL of cold 1 × PBS, centrifuged again at 100,000×g for 70 min, resuspended in 100 µL of 1 × PBS, and stored immediately at 80 ℃ (Aguiar et al. 2018 (link); Liu et al. 2018 (link)).
Transmission electron microscopy (TEM) was used to identify Exos. The Glaciostem transmission electron microscope (GLACIOSTEM, Thermo Fisher Scientific, USA) was adopted for observing and photographing.
Dynamic light scattering (DLS) was used to measure the diameter of Exos particles: the Exos sample was diluted to a ratio of 1:50 and the Zetasizer Ultra instrument (at Malvern Panalytical, UK) was used to detect the diameter of Exo when excited with a light wavelength of λ = 532 nm in a mixture of 0.15 M NaCl.
The BCA protein analysis kit (P0011, Shanghai Beyotime Biotechnology, China) was used to measure the protein concentration in Exos particles. Then, Western blot was applied to detect the expression of Exos-related proteins such as CD9, CD63, and TSG101.

The hydrodynamic diameter (d_H) and zeta potential of self-assembled nanostructures of CPT-S-S-PEG-CUR at various concentrations (0.25, 0.5, 1.0, and 2.0 mg/mL) in the presence and absence of TA (5 mg/mL) in an aqueous medium (Milli-Q water) were measured at 37 °C by using a Malvern Zetasizer Ultra instrument (Malvern, UK).