Cm100 tem

The MIM structure on Si was attached to a glass slide with a double-sided tape. Then, a solution of PVA (Carolina Biological Supply Company, molecular weight 45 g/mol) was poured onto the MIM structure so that the PVA flooded the entire Si substrate. The PVA was then left to dry in air overnight at room temperature. Then, the portion of the PVA film covering the MIM structure was excised with a razor blade and tweezers. The PVA film containing the MIM structures were dissolved in 200 µL of deionized water at ~70 °C until the film dissolved completely in water. The solution was then dropcast onto a Cu TEM grid with Carbon backing (Ted Pella) for TEM imaging. The TEM images were acquired with Phillips CM100 TEM with the operating voltage of 80 kV.
For in vitro imaging, 100 µL of the MIMs dispersed in water was incubated with T24T bladder cancer cells (American Type Culture Collection) for an hour. The cells were imaged with Zeiss LSM780 that is equipped with confocal upconversion fluorescence imaging using a tunable NIR Ti:Sapphire laser (Coherent Chameleon Ultra II).

Sections, approximately 80 nm thick, were cut with a Leica UC7 microtome and collected on copper grids with Formvar supporting membranes. Sections were stained with uranyl acetate and lead citrate and subsequently examined with a Philips CM 100 TEM (Philips, Eindhoven, The Netherlands), operated at an accelerating voltage of 80 kV. Digital images were recorded with an OSIS Veleta digital slow scan 2k x 2k CCD camera and the ITEM software package.

For transmission electron microscopy kidney samples were fixed using 4% PFA in PB by perfusion via A. renales using small silicone catheters. After perfusion fixation the kidneys were removed and sliced into 50 micrometer thin vibratome sections. Immunogold pre-embedding labeling was performed as described earlier^{47 (link)}. Briefly anti-CORO2B antibody was incubated overnight at 4 °C, sections were washed in PB and incubated overnight in secondary antibody at 4 °C (1.4 nm nanogold, Nanoprobes Inc., NY, USA). Gold labeling was enhanced using HQsilver kit (Nanoprobes Inc., NY, USA). Finally sections were embedded in Durcupan resin (Sigma-Aldrich, Germany) and ultrathin (40 nm) sections were cut using a Leica UC6 ultracut. Sections were imaged using a Philips CM 100 TEM.

HepG2 and Chang liver cell lines (1 × 10⁶ cells) were cultured in a completed medium with or without Hep88 mAb at IC50 concentration (12.5 μg/mL) for 0, 1, 2 and 3 days. The preparation procedures for the transmission electron microscope examination, including cell harvesting, glutaraldehyde/paraformaldehyde fixation, osmium post fixation, graded step alcohol dehydration and resin polymerization, were done as previously described [26 ]. Subsequently, 60–90 nm ultrathin sections were performed by a Porter-Blum MT-2 ultramicrotome and investigated under a Philips CM 100 TEM at 80 kV.

For transmission electron microscopic analysis, 2 μl of sample from fraction 6 of the sucrose density gradient was placed on a glow-discharged carbon-coated 400-mesh copper grid (CF400-Cu-50, Electron Microscopy Sciences, Hatfield, PA)) followed by negative staining with uranyl formate (1%, pH 6.8) for 30 s, and then examined at 80 kV using a transmission electron microscope (Philips CM100 TEM) equipped with CCD camera in the Imaging and Analysis Center (IAC) of Princeton University.

Scanning electron microscopy (SEM) was used to illustrate the morphology of the fiber mat samples. The SEM used was a Carl Zeiss EVO MA10 (White Plains, NY, USA) system with a 5–30 kV impinging voltage. Transmission electron microscopy (TEM) images of the samples were obtained using Phillips CM100 TEM (Nashville, TN, USA) under an accelerating voltage of 100 kV. The X-ray photoemission spectroscopy (XPS) spectra were obtained with a PHI Quantera SXM (Chanhassen, MN, USA) using monochromatic Al-Ka with an energy of 1486.6 eV to analyze the chemical composition of the surface. Raman spectra were collected using a He-Ne laser (wavelength of 633 nm and power of 17 mW) on a confocal Raman imaging system (Horiba Jobin Yvon LabRam Aramis, Edison, NJ, USA). Fourier transform infrared spectroscopy (FTIR) spectra of the fiber mats were obtained using the Spectrum 400 FT-IR spectrometer, PerkinElmer (Waltham, MA, USA).

Five µL of the AMP dispersion in Milli-Q water (25 µg/mL) containing 0.1% trehalose were added onto the glow discharge activated (30 s, 1 kV, 100 mA) [100 (link)] carbon/formvar coated 300 mesh copper grids for 30 s. The excess liquid was then blotted with a filter paper before the grids were air-dried at room temperature. The analysis was performed using the AnalySis 5.2 software suite (version 5.2; EMSIS, Germany) on images obtained by a Philips CM100 TEM (Philips EO, The Netherlands) instrument equipped with Veleta slow-scan CCD camera (EMSIS, Münster, Germany), at a magnification of 64,000×, giving a pixel size of 0.3 nm. All measurements were conducted at 80 kV. The statistics and size distribution graphical output were carried out in R (R Core Team).