Tecnai tf 20 feg tem

The cells were fixed in 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer pH7.4 for 1 h. They were then rinsed in 0.1 M sodium cacodylate buffer, scraped and pelleted in 2% agar. Samples were trimmed and post-fixed in 1% osmium tetroxide for 1 h, en bloc stained in 2% uranyl acetate in maleate buffer pH5.2 for a further hour, rinsed then dehydrated in an ethanol series and infiltrated with resin (Embed812, Electron Microscopy Science) and cured overnight at 60 C°. Hardened blocks were cut using a Leica UC7 Ultramicrotome, 60 nm sections were collected onto formvar/carbon coated nickel grids and stained using 2% uranyl acetate and lead citrate. These were viewed FEI Tecnai Biotwin TEM at 80 kV. Images were taken using Morada CCD and iTEM (Olympus) software typically at 26,000 × magnification. For electron tomography, 250 nm sections were collected on formvar/carbon copper grids, 10 nm gold particles added on both sides of the grids (Utrect UMC). A tomography tilt series was acquired using SerialEM software on an FEI Tecnai TF20 FEG TEM at 200 kV. tomograms were reconstructed using IMOD software (University of Colorado, Boulder, CO).

The structure and morphology of the prepared LM nanocapsules were observed using
high-resolution TEM (EM-002B; Topcon, Tokyo, Japan) with an accelerating voltage
of 120 kV. A small droplet of sample placed on a grid or
10 μl of sample solution (LM concentration:
100 μg ml⁻¹) were
irradiated using a fibre-coupled CW laser at 785 nm for
3 min (spot diameter, ∼4 mm; maximum power:
1 W,
∼80 mW mm⁻²;
BRM-785-1.0-100-0.22-SMA; B&W Tek, Newark, DE, USA). The polymer shell
structure of nanocapsules and STEM/EDS mapping were performed by Nanoscience Co.
in Evans Analytical Group Company, Inc. (Tokyo, Japan). The samples were imaged
with a FEI Tecnai TF-20 FEG/TEM operated at 200 kV in bright-field
TEM mode, high-resolution (HR) TEM mode, and high-angle annular dark-field
(HAADF) STEM mode. The STEM probe size was 1–2 nm nominal
diameter. EDS mapping were acquired on Oxford INCA, Bruker Quantax EDS
system.
The hydrodynamic diameter of LM nanocapsules was examined by DLS (Photal
FPAR-1,000; Otsuka Electronics, Osaka, Japan). DLS diagram of laser-induced LM
nanocapsules was also measured. A 100 μl of sample solution
(LM concentration:
100 μg ml⁻¹) was
irradiated using a fibre-coupled CW laser (maximum power: 1 W,
∼80 mW mm⁻²) at
785 nm for 1 h before DLS measurements.
The concentration of LM and carmofur in nanocapsules was estimated with a
ultraviolet–visible–NIR spectrophotometer (V-730 BIO; Jasco,
Tokyo, Japan).

In order to observe the deformation process beneath the indenter tip, as well as the shear plasticity, a bright field cross sectional transmission electron microscopy (XTEM) images are carried out on selected indents using FEI Tecnai TF-20 FEG/TEM operated at 200 kV in bright-field (BF). The TEM-ready sample was prepared using the in-situ FIB lift out technique on a FEI Dual Beam FIB/SEM. The sample was capped with sputtered C and e-Pt/I-Pt prior to milling to protect the surface of the area of interest.