Samples of freshly collected aquaria biofilms and microbial mat samples were briefly rinsed in abundant distilled water to avoid the formation of extracellular precipitates upon drying, as previously described (Couradeau et al., 2012 (link), 2013 (link)). Then, small sample fragments were deposited onto on formvar-coated transmission electron microscopy (TEM) cupper microscopy grids and let dry. Scanning electron microscopy (SEM) analyses were performed using a Zeiss ultra 55 SEM equipped with a field emission gun. Images were collected in backscattered electron (BSE) mode with a Zeiss Ultra 55 FEG-SEM operating at 10 kV with a 30 μm aperture and a working distance of 7.5 mm using the Angle selective Backscattered (AsB) detector. This mode of imaging provides a contrast which is sensitive to the average atomic number, hence allowing detecting intracellular carbonate inclusions. Elemental compositions of the observed mineral precipitates were directly determined by energy dispersive x-ray spectrometry (EDXS) using an EDS QUANTAX detector and the software ESPRIT (Bruker Corporation, Germany) as previously achieved by Couradeau et al. (2013 (link)).
Esprit
Manufactured by Bruker
Sourced in Germany
Esprit is a versatile energy dispersive X-ray spectroscopy (EDS) system designed for materials analysis. It provides elemental identification and quantification capabilities for a wide range of sample types. The core function of Esprit is to perform high-quality, rapid, and precise elemental analysis.
Automatically generated - may contain errors
Lab products found in correlation
Ultra 55 sem, by Zeiss (1 mentions)
Ultra 55 feg sem, by Zeiss (1 mentions)
Xflash 4010, by Bruker (1 mentions)
Titan g2 80 200, by Thermo Fisher Scientific (1 mentions)
Quantax 400, by Bruker (1 mentions)
Fe sem, by Zeiss (1 mentions)
Quantax 800, by Bruker (1 mentions)
Avanti j e, by Beckman Coulter (1 mentions)
7 protocols using esprit
1
Scanning Electron Microscopy Analysis of Aquaria Biofilms
2
Multimodal SEM Imaging and FIB-TEM Sample Preparation
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We used a Zeiss Ultra 55 field emission gun SEM operated at 2 to 15 kV at IMPMC, Paris. Backscattered electron (BSE) mode was used to investigate chemical heterogeneities using an Angle Selective Backscattered Detector (AsB, working distance 7.5 mm) or an energy selective backscattered detector (EsB). Morphology imaging was performed using an InLens detector (working distance 2–3 mm). Energy dispersive X-ray spectrometry (EDXS) maps were acquired using an EDXS QUANTAX system equipped with a silicon drift detector XFlash 4010 (Bruker). Data were processed with the software Esprit (Bruker). Focused ion beam (FIB) milling was used to produce ultra-thin sample sections (Fig. 5d ) on a Zeiss neon EesB40 FIB/FEG-SEM system (IMPMC, Paris). A FIB-assisted Pt deposit was first made. A 30 kV Ga+ beam operated at ca 5 nA was then used for the initial milling steps, consisting in rough excavations from both sides of the thin foil. An in situ micromanipulator was attached to the foil by FIB-assisted platinum deposition before separation of the foil (at ca 100 pA). The thin foil was transferred to a TEM grid and welded to it. The thinning of the ultra-thin foil was performed with the beam operated at a ca 100 pA current. A last cleaning step was performed at low acceleration tension (ca 3 kV).
3
Multimodal TEM Analysis of Nanomaterials
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High-angle annular dark-field (HAADF) scanning TEM (STEM) imaging, energy-dispersive X-ray spectroscopy (EDXS) mapping and electron to-mography were performed in an FEI Titan G2 80-200 transmission electron microscope equipped with a high brightness field emission gun, a probe aberration corrector and an in-column Super-X EDXS system. HAADF STEM images were recorded on a Fischione detector using a beam convergence semi-angle of 24.7 mrad and an inner detector semi-angle of 69 mrad. The chemical compositions were analyzed using Esprit (Bruker) software.
4
Elemental Mapping via SEM-EDX Analysis
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Energy-dispersive X-ray (EDX) elemental mappings were acquired in a JEOL JSM-7401F SEM at 20 kV using a Quantax 400 (Bruker, Billerica, MA, USA) energy-dispersive X-ray spectrometer. The working distance was set to ~12.0 mm. At least three different areas were analyzed via EDX analysis for each sample. The measurement duration was set to at least 300 s. For the evaluation of the acquired spectra, the software Esprit (Bruker, V1.8.2 (2008)) was used.
5
STEM-EDX Mapping with FEI Titan G2
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A FEI Titan G2 80-200 is used for the experiment of STEM-EDX mapping, equipped with Schottky field emission electron gun (working at 80 kV), a Cs probe corrector (DCOR, CEOS), and a Super-X EDX system with four detectors. Cliff-Lorimer method is applied for the quantification of specimens in the EDS map-analysis with build-in software (Esprit, Bruker).
6
Comparative Analysis of XRF Scanners
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The areas scanned by both the Modular Scanner and the M4 Tornado are highlighted in Figure 1 . Although M4 Tornado can provide considerably shorter dwell-times, thanks to its higher power X-ray tube, both analyses were performed under the most similar conditions possible, to ascertain whether the results are comparable. Scanned areas were 5.3 × 5.0 cm in size each and were acquired with a tube voltage of 35 kV and with an applied current of 29 μA. The areas scanned with each instrument were slightly offset from one another and were cropped to improve the comparison and overlay between them.
Dwell-times were set to 500 ms for the M4 Tornado and 470 ms for the Modular Scanner, with an overall stage speed of approximately 2 mm/s. The M4 Tornado scanner is equipped with a Rh target X-ray tube, while the Modular Scanner had an Ag target Moxtek® tube attached. The Modular Scanner was assembled with the mobile stage and two X-123 SDD AMPTEK® detectors.
Overall scanning time was about 20 min for each instrument, with both acquisitions being performed in a continuous scan fashion. Elemental distribution maps were generated by different software. M4 Tornado images were obtained with Bruker ESPRIT proprietary software. Data obtained with the Modular Scanner were processed with an in-house developed software (XISMuS) version 1.3.2 [21 (link)].
Dwell-times were set to 500 ms for the M4 Tornado and 470 ms for the Modular Scanner, with an overall stage speed of approximately 2 mm/s. The M4 Tornado scanner is equipped with a Rh target X-ray tube, while the Modular Scanner had an Ag target Moxtek® tube attached. The Modular Scanner was assembled with the mobile stage and two X-123 SDD AMPTEK® detectors.
Overall scanning time was about 20 min for each instrument, with both acquisitions being performed in a continuous scan fashion. Elemental distribution maps were generated by different software. M4 Tornado images were obtained with Bruker ESPRIT proprietary software. Data obtained with the Modular Scanner were processed with an in-house developed software (XISMuS) version 1.3.2 [21 (link)].
7
Visualizing Toothpaste Abrasives
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For visual observation of the toothpaste abrasives, 1 g of toothpaste and 50 mL of distilled water were thoroughly stirred until completely dissolved. The solid was centrifuged at 10,000 RPM for 15 min using a High-Speed Centrifuge (Avanti J-E, Beckman Coulter, Fullerton, CA, USA). The supernatant was discarded. The process was repeated five times by adding fresh distilled water. Finally, the solids were washed out with ethanol to remove residual components. The remaining solid was dried for 3 days in a dryer heated to 37°C. The RS was used as calcium pyrophosphate powder without washing process. The powders were attached to the stub with copper tape and coated with platinum particles. The image and composition of the powders were investigated using FE-SEM (AURIGA, Carl Zeiss, Jena, Germany) and EDS (QUANTAX 800, Bruker, Berlin, Germany) with analysis software (ESPRIT, ver. 2.1.2.17832, Bruker). All groups were operated and magnified at 11 kV and 1,000×. Additionally, R2 was magnified at 500×.
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