Lext ols3000

To compare the sections prepared by ultramicrotomy and the TEM lamellas prepared by in situ FIB lift-out, as well as to demonstrate specific preparation artefacts, a further microscopic examination was performed. The data used to evaluate TEM sample preparation were obtained by scanning electron microscopy (Quanta FEG250; FEI, Frankfurt am Main, Germany) at a primary beam energy ranging from 5 to 10 keV with a large field detector (LFD). Surface morphology was examined using reflected-light microscopy (LEXT OLS 3000; Olympus, Hamburg, Germany), confocal laser scanning microscopy (LEXT OLS 3000; Olympus, Hamburg, Germany) and atomic force microscopy (Nanowizard I^®, JPK-Instruments, Berlin, Germany). High-resolution AFM images were generated in contact mode using a standard CSC37 cantilever (Ultrasharp CSC37/ no AL, MicroMasch, Tallin, Estonia) with the following parameter settings: cut-off frequency, 150 Hz; amplification factor, 0.05; scan size from 100 µm × 100 µm to 2 µm × 2 µm; scan rate, 0.5 Hz; setpoint, 0.5 V at Vsum = 1.5 V; resolution, 512 × 512. AFM measurements were performed using JPK data-processing software (v.5.0.97).

Surface topography was investigated on the specimens of each subgroup using a three-dimensional (3-D) confocal laser scanning microscopy (CLSM; LEXT OLS3000, Olympus, Tokyo, Japan) at 50× magnification. The areal texture parameters were obtained using a software (LEXT-OLS, version 6.0.3, Olympus, Tokyo, Japan): Sa, the arithmetic mean height; Sq, the root mean square height; and Sv, the maximum pit height of the scale-limited surface according to ISO 25178 [20 (link)]. Surface measurements were processed with the form and outlier eliminated. Tilt was corrected and a 3-D surface was constructed with the distance to the optical center in the X axis, the tilt angle on the Y axis, and the flatness error on the Z axis. A robust short wavelength pass Gaussian filter (cut-off wavelength: 10 μm) was applied to the data in order to decompose waviness from roughness. For each specimen, three different measurements (effective field of view was 256 × 192 μm) on the either polished sides for controls or sandblasted sides for experimental subgroups were performed. A total of 30 measurements was obtained for each subgroup.

The density of the rubbery filler was measured using a pycnometer in acetone. The density of porous matrices was obtained directly from the weight (8–18 g, by analytical balance) divided by the volume of the sample corresponding to the volume difference of ethanol added to a narrow-necked container in the presence/absence of the sample (measurement repeated with three samples) [32 (link)].
The porosity of the matrix leading to the calculation of the hypothetical density of the non-porous matrix was determined using confocal laser scanning microscopy (Lext OLS 3000, Olympus). The value used was the average of 10 measurements. The measurement was performed as the rate analysis of void cross-section sum in fracture area in optical mode [32 (link)].
The tensile test was performed using a universal static material testing machine (ZWICK Z010 ROELL). The strain rate was 30 mm∙min⁻¹. Tensile modulus (E), ultimate strength (σ_Fmax), ultimate strain (ε_Fmax), and specific energy needed for ultimate strength achievement (A_Fmax) were determined from the measurement. The tensile modulus was determined from the linear part of the tensile curve in the strain range of 0.05–0.25 % [32 (link)]. Each sample series included five tested samples to achieve good reproducibility of results.