Ht7700 microscope

Exosomes in culture medium of CL187 cells were isolated by differential centrifugation. Briefly, cells were grown in high‐glucose DMEM supplemented with 10% exosome‐depleted FBS. After 48 hours, culture medium was collected and subjected to sequential centrifugation at 300 g for 5 minutes and 15,000 g for 15 minutes at 4°C to remove residual cells and debris. After filtered with a 0.22 μm Millex‐GV filter (Millipore, Billerica, MA, USA), the resultant supernatant was ultracentrifuged at 150,000 g for 3 hours at 4°C, and the exosome pellets were resuspended in PBS for use. For the analysis of plasma‐derived exosomes, peripheral blood samples (4 mL each) were collected in anticoagulant tubes from healthy donors or CRC patients, and the supernatant was obtained by centrifugation at 2000 g for 10 minutes. The exosomes were isolated by ultracentrifugation as above.
The morphologic features of exosomes were characterized by negative staining electron microscopy. The images were taken by a transmission electron microscope (HT7700 Hitachi microscope, Tokyo, Japan) at 100 kV. Two exosome markers, TSG101 and CD63, and LEA were detected by western blotting.

UV–Vis absorption spectra were recorded using a UV-1900 spectrophotometer (Shimadzu). Fluorescence spectra were recorded using a FluoroMax-4 (Horiba). CD was characterized by JASCO-1500 (JASCO). CPL spectra were recorded using a ChirascanV100 Circular Dichroism Spectrometer (Applied Photophysics). TEM characterization was performed on a Hitachi HT7700 microscope operating at 100 kV. AFM characterization was performed on a Bioscope Resolve Atomic Force Microscopy (Bruker).

The crystal structure of the Pt-based samples was characterized by X-ray powder diffraction (XRD) using a Rigaku D/max-ga X-ray diffractometer with graphite monochromatized Cu Kα radiation (λ = 1.54178 Å). Transmission electron microscopy (TEM) images of such samples were taken using a HITACHI HT-7700 microscope operated at 100 kV. High-resolution transmission electron microscopy (HRTEM) was performed using a FEI Tecnai F30 G2 microscope operated at 300 kV. High-angle annular dark-field scanning TEM (HAADF-STEM) and Energy dispersive X-ray (EDX) mapping analyses were taken on a FEI Titan ChemiSTEM equipped with a probe-corrector and a Super-X EDX detector system and operated at 200 kV. The percentages of the elements in the samples were determined using inductively coupled plasma atomic emission spectrometry (ICP-AES, IRIS Intrepid II XSP, TJA Co., USA). Gas chromatography mass spectrometer (GC-MS) measurements were performed on a GC-MS 7890A-5975C (Agilent) with molecular ion selective monitoring. All of these samples were diluted with acetone in fixed ratio before the GC-MS measurement.

Transmission electron microscopy (TEM) images were acquired using an HT-7700 microscope (Hitachi, Tokyo, Japan) operated at 75 kV to investigate the morphologies of the fabricated NPs. The TEM images of the hollow NPs were used to measure their outer and inner diameters. Scanning electron microscopy (SEM) images were acquired using an SU-8220 microscope (Hitachi, Tokyo, Japan) operated at an accelerating voltage of 3 kV to determine the outer diameters of the hollow NPs and the sizes of their openings. The mean values were calculated using the results of over 100 particles randomly selected from each sample, and the standard deviation was represented by the errors. The ζ-potential and size distribution of the NPs were investigated using a Zetasizer Nano-ZS dynamic light-scattering (DLS) analyzer (Malvern Instruments, Worcestershire, UK). In order to demonstrate the size distribution, the standard deviation was represented by the errors. The inclusion of the FA mixture in the NPs was confirmed using differential scanning calorimetry (DSC) (Q2000, TA Instrument, New Castle, DE, USA). All the measurements were performed in the range of 20 to 230 °C at a scanning rate of 2 °C/min.

Both the morphology and ultrastructure of the ZJU-0430 cells were examined following seeding into 25-mm³ tissue-culture flasks. The general morphology of the cells was observed daily under a phase-contrast microscope and histopathologically compared to that of the original tumours. Electron microscopy was conducted as previously described [32 (link)] using the GBC-SD cell line as the control. Cells were harvested, pelleted, fixed (2.5% glutaraldehyde), post-fixing (2% osmium tetraoxide) and then embedded in EPON resin. Ultrathin sections were stained and examined under a HT7700 microscope (Hitachi, Tokyo, Japan).

The surface morphology of AC-NCDs was characterized by using the transmission electron microscopy (TEM; Hitachi HT-7700 microscope). The functional groups of AC-NCDs were recorded by performing FTIR spectroscopy (ALPHA FTIR Spectrometer, Bruker). The UV-vis absorbance was measured by using Spectra Academy UV–Visible Spectrometer Detector SV-2100. Raman spectrum of AC-NCDs was measured by Micro Raman Identify Spectrometer from ProTrusTech Co. Ltd. (Tainan, Taiwan). The fluorescence spectra were monitored with a Varian Cary Eclipse Fluorescence Spectrophotometer. The zeta potential values were done by using a NanoPlus HD-Zeta/Nano Particle Analyzer.

10 μL ADSC-EVs (the concentration of ADSC-EVs used in this experiment was 20 μg/mL) were first added to a formvar-carbon-coated grid. After ADSC-EVs had been allowed to be adhered to the grid for 30 min, the grid was rinsed with droplets of deionized water and fixed for 5 min with 1% glutaraldehyde. The sample was then stained with 2% uranyl acetate. After removing remnant liquid using a blotting paper, the grid was dried at room temperature. Photographs were taken using a HT7700 microscope (Hitachi, Tokyo, Japan) at an acceleration voltage of 80 kV.