Utricles were fixed with 2.5% glutaraldehyde (Electron Microscopy Sciences) in 0.1 M phosphate buffer (PB) at 4 °C overnight, post-fixed with 1% aqueous OsO4, dehydrated in a graded ethanol series, and dried by critical point drying with liquid CO2 (EM CPD300; Leica). Specimens were coated with 100 Å Au using an E-1045 sputter coater (Hitachi, Tokyo, Japan), and analysed with a NOVA NanoSEM 230 scanning electron microscope (FEI, Hillsboro, Oregon, USA) operated under a high vacuum at 5–10 kV at a working distance of 6–7 mm. The orientation of individual hair cells was measured using the ImageJ (NIH, USA) angle measurement tool. As outlined in Fig. 4A , stereocilia bundle polarity was measured in five analysis fields positioned in the posterior striola (PS), medial striola (MS), anterior striola (AS), medial extrastriola (MES), and lateral extrastriola (LES) separately. The detailed angle calculation method was performed as Deans described previously29 (link). Vestibular hair cell orientation was assembled as a circular histogram using Matlab 2014a (Mathworks, USA) software.
Nova nanosem 230 scanning electron microscope
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Nova NanoSEM 230 is a scanning electron microscope (SEM) designed for high-resolution imaging of a wide range of samples. It features a field emission gun (FEG) electron source and advanced electron optics to provide high-quality, low-noise images at magnifications up to 1,000,000x. The Nova NanoSEM 230 is capable of operating in high and low vacuum modes, allowing for the examination of both conductive and non-conductive samples.
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Lab products found in correlation
E 1045 sputter coater, by Hitachi (2 mentions)
Em cpd300, by Leica (2 mentions)
Matlab 2014a, by MathWorks (1 mentions)
Glutaraldehyde, by Electron Microscopy Sciences (1 mentions)
Axis ultra dld x ray photoelectron spectroscopy, by Shimadzu (1 mentions)
Tecnai g2 f20 transmission electron microscope, by Thermo Fisher Scientific (1 mentions)
D8 advance x ray diffractometer, by Bruker (1 mentions)
Asap 2020 physisorption analyzer, by Micromeritics (1 mentions)
Energy dispersive x ray spectrometer, by Thermo Fisher Scientific (1 mentions)
4294a impedance analyzer, by Agilent Technologies (1 mentions)
A1 confocal laser microscope, by Nikon (1 mentions)
Nis elements software, by Nikon (1 mentions)
7 protocols using nova nanosem 230 scanning electron microscope
1
Scanning Electron Microscopy Analysis of Vestibular Hair Cell Orientation
2
Comprehensive Characterization of Materials
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X-ray powder diffraction (XRD) patterns were measured by a D8 ADVANCE X-ray diffractometer (Bruker, Germany) with Cu Kα radiation. The C content was measured by vario EL cube elemental analysis system (Elementar, Germany). The morphologies of products and EDX spectra were performed using a Nova Nano SEM 230 scanning electron microscope (FEI, USA) and a Tecnai G2 F20 transmission electron microscope (FEI, USA). X-ray photoelectron spectroscopy (XPS) was obtained by an AXIS Ultra DLD X-ray photoelectron spectroscopy (Shimadzu, Japan). N2 adsorption–desorption isotherms were measured by an ASAP 2020 Physisorption Analyzer (Micromeritics, USA), and the specific surface areas were calculated by the Brunauer–Emmett–Teller (BET) method.
3
Microstructure Analysis of 3Y-TZP/Ta and Titanium Alloy
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The microstructure of the polished 3Y-TZP/Ta and titanium alloy disk specimens was studied using a Nova NANOSEM 230 scanning electron microscope (SEM, FEI, Hillsboro, OR, USA). A 3D surface Talysurf CLI 500 profilometer (Taylor Hobson, Leicester, UK) was used to measure the roughness ratio or specific surface area (the ratio between the actual surface and projected area, Sdr) and the average surface roughness (Ra) of the samples by scanning the surface with a stylus. Data are presented as the mean value with the corresponding error of ten independent experiments. The stylus arm had a 90° diamond tip with a nominal radius of 2 μm. The data sampling intervals in X and Y were 0.5 and 2.5 μm, respectively. The Z-scale resolution was 32 nm. The profilometer generated a 3D map of the surface topography. The disks were rinsed with sterile saline to remove loosely attached cells after biofilm formation and then air-dried and examined by SEM.
4
Orai1 Deficiency Impacts Bone Microstructure
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The analysis was performed at UCLA department of materials science and engineering core facility using a Nova NanoSEM 230 scanning electron microscope (FEI, USA) with field emission gun and variable pressure capabilities equipped with backscattered electron detectors and an energy dispersive x-ray spectrometer (ThermoScientific, USA). Femur samples from 8-week old Orai1−/− male mice and sex-matched WT littermate were fixed in 4% Glutaraldehyde overnight at 4′C, non-decalcified, resin-casted and coronal-sectioned at the distal metaphyseal area, sputter-coated with gold palladium, and subsequently examined with SEM.
5
Characterization of PUEH and PZT Transducers
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For taking the optical microscope images, a DFC290 optical microscope (Leica) is used. For taking the SEM images, a Nova NanoSEM 230 scanning electron microscope (FEI Company) is used. For the impedance measurements of the PUEH and bulk PZT transducer, a 4294A impedance analyzer (Agilent) is used. A 33500B signal generator (Agilent) is used to generate sinusoidal signals to bulk PZT transmitter. For voltage measurement, the output signal is connected to a DSO-X3034A oscilloscope (Agilent).
6
Scanning Electron Microscopy of Utricles
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Utricles were fixed with 2.5% glutaraldehyde at 4°C overnight and then processed using 2% tannin and 1% osmium acid. Graded ethanol series were used to dehydrate the samples, and liquid CO2 (EM CPD300; Leica, Wetzlar, Germany) was used to dry them. An E-1045 sputter coater (Hitachi, Tokyo, Japan) was used to coat specimens with 100 Å Au. A NOV A NanoSEM 230 scanning electron microscope (FEI, Hillsboro, OR, USA) was used to scan the samples.
7
3D Collagen Scaffold Characterization
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Cells seeded in 3D collagen scaffolds were imaged by SEM and Laser Confocal Microscopy as previously described50 (link). For SEM imaging samples were washed 3 times with 0.1 M sodium cacodylate buffer pH 7.4, fixed in 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer pH 7.4 for 2 h at 4 °C and washed again in 0.1 M sodium cacodylate buffer pH 7.4 (Sigma Aldrich). Samples were then dehydrated in a graded series of ethanol for 10 min each, dried in a dessicator overnight and sputter-coated with platinum. Images were acquired with a Nova NanoSEM 230 scanning electron microscope (FEI, Hillsboro, OR, USA). For confocal imaging, cells were washed 3 times with 1% PBS, fixed with 4% paraformaldehyde for 20 minutes at room temperature and stained with 10 µM/ml DRAQ5™ (ImmunoChemistry Technology, Bloomington, MN, USA). Images were acquired with an A1 laser confocal microscope (Nikon Corporation, Tokyo, Japan) and analyzed with the NIS Elements software (Nikon Corporation, Tokyo, Japan).
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