Scios dualbeam

Structural characterizations of Fe₃GeTe₂ and CrSb samples were carried out by X-ray diffraction (Bruker D8 Discover, Bruker Inc., Billerica, MA, USA) and TEM (FEI Tecnai F20) equipped with EDS. Sample composition and doping concentration were determined by EDS. Cross-section TEM samples were prepared by Focused ion beam (FEI Scios DualBeam).

A focused ion beam (FEI Scios Dual-beam) set to 5 nA at 30 kV was used to cleave off the end of the probe and expose the electrical and optical channels, revealing a final probe diameter of 8–12 µm for the microfiber cores. Electrochemical impedance spectroscopy (EIS) was carried out with the Versastat4 (running VersaStudio v.2.60.6) to determine probe impedance in a 1× phosphate-buffered saline (PBS) using the same reference and counter electrodes described above. Optical coupling efficiency was determined by measuring light output from a fiber optic patch cable (Thorlabs, P1405B-FC-5) using three light sources (473, 543, and 673 nm) interchangeably coupled into the cable. Light power was measured by placing the ferrule 5–10 mm away from the detector head of a digital power meter (Thorlabs, PM100D). A ceramic ferrule sleeve (Thorlabs, ADAL1) was then slid halfway onto the patch cable, and different EO-Flex probes were slid into the opposite end to couple light through. Light power from the tip of the EO-Flex probes was measured using a similar protocol to the patch cable.

Microliths were collected without dispase treatment from 20- to 28-week-old male Npt2b^−/− mice and a PAM patient explant as described above. Microliths were incubated with distilled water or 0.02% SDS for 5 min, then washed 5 times with water to remove residual SDS and dried. The samples were placed on conductive carbon tape. Imaging was performed at 2 kV on a scanning electron microscope (FESEM, FEI Scios DualBeam, Germany). The elemental analyses were obtained at 15 kV beam voltage using an EDAX Octane Elite Super detector.

FIB-SEM analysis was performed at the Institute of Geochemistry, Chinese Academy of Sciences, using a FEI Scios Dual beam that combines a traditional Field Emission electron column with a FIB column equipped with EDS, ETD (secondary electron, SE), and T1 (backscattered electron, BSE) detectors. In situ ion-milled lift-out procedures were performed by FIB-SEM to prepare ultrathin foils (<100 nm thick) for scanning transmission electron microscopy observation^{71 (link)}. Ultrathin foils of the filamentous microfossils were then observed under a FEI G2 F20 S-TWIN transmission electron microscope (at 200 keV) at the State Key Laboratory of Mineral Deposits Research, Nanjing University. Elemental compositions of the microfossils and matrix were detected on an OXFORD EDS X-Max 80 T instrument. Scanning electron microscopy (SEM) imaging was performed using a Phenom ProX equipped with BSE and EDS detectors.

The resonators were built in the following way. After pre-indenting a 200 µm rhenium gasket to about 15–20 µm, a thin layer of copper (1–2 µm) was deposited on the diamonds. The shape of the LLs was cut out from the copper layer using a focused ion beam (Scios Dual beam from FEI), Fig. 2b. As a result, the first stage LL typically runs from the outer rim of the diamonds pavilion toward close to the rim of the diamonds culet, with a thin 15 µm slit running all along the 1 mm pavilion (Fig. 2b). The second stage LL, is typically placed on the culet face having about 230 µm outer diameter and 80 µm inner diameter which closely follows the geometry of the gasket hole.
To ensure electrical insulation between both LLs and the metallic gasket, a 1 µm layer of Al₂O₃ was deposited on the gaskets.

Miniaturized specimens for the push-out tests were produced using a FIB (FEI Scios Dual beam, Oregon, USA) milling technique. The milling process is illustrated in Figure 2. First, a rectangular volume was milled out by applying the ion beam perpendicular to the top surface and close to the polished edge of the coupon, as shown in Figure 2a. A milling current of 5 nA was used in this step. A rectangular Au/SiN bi-layer panel attached to the GaAs base was created. A trench was then milled into the GaAs substrate by positioning the ion beam at a right angle to the polished cross-section, as shown in Figure 2b. The milling current used was 5 nA. As shown in Figure 2c,d, the inner face at the deeper end of the trench was parallel to the SiN/GaAs interface and a very thin sheet of GaAs was left attached to the SiN film. The thickness of the GaAs sheet is in the range of 60–80 nm. Five specimens in total were fabricated. SEM images of a typical push-out specimen are shown in Figure 3. The depth (h_t), length (L_t), and width (W_t) of the trench, as well as the length (L_p) and width (W_p) of the rectangular Au/SiN panel, were measured using SEM, as given in Table 2.

We further assessed the presence or absence of melanosome hollowness using transmission electron microscopy (TEM) for extant species (in which this was not documented elsewhere, Fig. S12) and focused ion beam sectioning (FIB‐SEM) for fossil samples. Briefly, for TEM analyses, feathers were embedded in resin following the protocol from Shawkey et al. (2003), see Supporting Information Methods for details), and then cut into 70–100 nm thick cross sections with an RMC‐MT ultramicrotome 6000. Cross sections were placed on copper grids and observed with an FEI Morgagni 268D transmission electron microscope. FIB‐SEM methods follow those described in Vitek et al. (2013) and Schiffbauer and Xiao (2009, 2011), using an FEI Scios DualBeam at the University of Missouri Electron Microscopy Core Facility. FIB‐SEM milling was conducted with a Ga⁺ ion beam voltage at 10 kV. Overview SEM images following FIB‐SEM milling were collected using a Zeiss Sigma 500 VP at the University of Missouri X‐ray Microanalysis Core Facility.