Surface hardness was characterized by Vickers-hardness and Nano-indentation hardness. The Vickers-hardness measurements were measured using a micro-hardness instrument (MVS 1000D1, Guangjing, Guangzhou, China) with a load of 0.98 N, dwell time of 15 s; The Nano-indentation measurements were carried out using a Nano-indentation instrument (G200, Keysight, Santa Rosa, CA, USA) with diamond Berkovich indenter, which selected the dynamic nano-indentation model with the maximum indentation depth of 2500 nm. Three measured data for each specimen were averaged to reduce errors caused by different positions. To reveal the microstructure, samples were etched with a 50 mL H2O, 25 mL HNO3 and 5 mL HF solution. The cross-sectional microstructure was observed by optical microscope (DMI 5000 M, LEICA, Weitzlal, Germany) and transmission electron microscope (TECNAI G2 S-TWIN F20, FEI, Hillsboro, OR, USA). Thin TEM foils were prepared via an ion-miller (PIPS-691, Gatan, Pleasanton, CA, USA) with a beam voltage of 3.5 kV and a milling angle of 4–8°. TEM images and the corresponding selected area electron diffraction (SAED) were obtained by TEM with an accelerating voltage of 200 kV.
Dmi5000m
Manufactured by Leica camera
Sourced in Germany
The DMI5000M is an inverted microscope designed for advanced microscopy applications. It features high-performance optics, precise motorized controls, and a modular design to accommodate a variety of advanced imaging techniques. The DMI5000M is a versatile tool for researchers and scientists working in fields such as cell biology, materials science, and more.
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Lab products found in correlation
Pips 691, by Ametek (1 mentions)
Tecnai g2 f20 s twin, by Thermo Fisher Scientific (1 mentions)
3 diaminobenzidine, by Solarbio (1 mentions)
Tegrapol 35, by Struers (1 mentions)
Mht 10 microhardness tester, by Anton Paar (1 mentions)
Supra 40, by Zeiss (1 mentions)
X pert pro mpd, by Malvern Panalytical (1 mentions)
Highscore plus, by Malvern Panalytical (1 mentions)
Dm6000b, by Leica camera (1 mentions)
Dsc404, by Netzsch (1 mentions)
Biotin conjugated affinipure goat anti rabbit igg h l, by Proteintech (1 mentions)
Alizarin red, by Beyotime (1 mentions)
Application suite, by Leica camera (1 mentions)
Dropshape analysis, by Krüss (1 mentions)
Easydrop instrument, by Krüss (1 mentions)
X pert pro mrd, by Malvern Panalytical (1 mentions)
11 protocols using dmi5000m
1
Characterization of Surface Hardness
2
Immunohistochemical Analysis of MAP2 Expressing Neurons
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Brain sections were rinsed with PBS, treated with 0.3% H2O2 for 15 min, and rinsed in PBS (three times for 5 min each). Sections were then blocked with 1% goat serum albumin (SL039; Solarbio, Beijing, China) for 10 min at room temperature and incubated overnight at 4 °C with a MAP2 primary antibody (17490-1-AP, 1:100; Proteintech, Rosemont, IL, USA). Sections were rinsed and incubated in the appropriate secondary antibody for 1 h at room temperature. A solution containing 3′-diaminobenzidine (Solarbio, Beijing, China) was added after incubation with the secondary antibodies. Sections were then dehydrated using an alcohol gradient, cleared in xylene, and mounted for microscopic examination (DMI5000M; Leica) to count the number of positive cells.
3
Tensile Specimen Microstructural Characterization
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Microstructural characterization of the tensile specimens was performed by optical microscopy using Leica DMI 5000 M and by SEM. A horizontal and a vertical plane from a Z tensile specimen were polished on a Struers TegraPol-35. The samples were polished with a final sand paper grit size of 1200 grit and subsequently with diamond suspension from 6 µm to 1 µm sizes. The microsections were then etched with a Ti ASTM 186 chemical agent for 30 s.
The Vickers Hardness (HV) was measured on the microsections of tensile specimens produced with both powder batches, on one horizontal (X orientation) sample per batch and one vertical (Z orientation) sample per batch, i.e., on a total of four samples. HV was measured using an MHT-10 Microhardness tester (Anton Paar GmbH, Graz, Austria), using a load of 200 gf with a 30 s hold time, and measuring with a 50× optical magnification.
The Vickers Hardness (HV) was measured on the microsections of tensile specimens produced with both powder batches, on one horizontal (
4
Quantifying Colonic CD11c+ Cells
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Fixed colonic samples were cut into 4-μm-thick sections, deparaffinized and rehydrated through a series of xylene and ethanol washes. Sections were blocked with 1% goat serum albumin for 10 min at room temperature and incubated overnight at 4 ℃ with an CD11c primary antibody (Abcam, Cambridge, UK). Then, a horseradish peroxidase‑conjugated secondary antibody was added and incubated for 60 min at 37 ℃. The sections were then dehydrated using an alcohol gradient, cleared in xylene and mounted for microscopic examination (DMI5000M; Leica, Solms, Germany) to count the number of positive cells.
5
Comprehensive Corrosion Product Analysis
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The morphology
and composition of the corrosion products were observed with scanning
electron microscopy (SEM, ZEISS SUPRA 40, Germany) with 10 kV acceleration
voltage and energy dispersive spectroscopy (EDS, AZtec, Oxford, UK).
The chemical composition of the corrosion products was analyzed by
means of X-ray diffraction (XRD, X’pert Pro MPD Panalytical,
Netherlands), with monochromated Cu-Ka radiation at the 2θ range
of 10–80°. The software of HighScore Plus (Panalytical,
Almelo, Netherlands) was selected to analyze the data. An optical
microscope was used to measure the depth of corrosion pits on the
steel surface after removing the surface corrosion products (DMI 5000M,
Leica).
and composition of the corrosion products were observed with scanning
electron microscopy (SEM, ZEISS SUPRA 40, Germany) with 10 kV acceleration
voltage and energy dispersive spectroscopy (EDS, AZtec, Oxford, UK).
The chemical composition of the corrosion products was analyzed by
means of X-ray diffraction (XRD, X’pert Pro MPD Panalytical,
Netherlands), with monochromated Cu-Ka radiation at the 2θ range
of 10–80°. The software of HighScore Plus (Panalytical,
Almelo, Netherlands) was selected to analyze the data. An optical
microscope was used to measure the depth of corrosion pits on the
steel surface after removing the surface corrosion products (DMI 5000M,
Leica).
6
High-Speed Particle Tracking and Analysis
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Measurements of 2.8 μm diameter particles were performed with a widefield upright microscope (Leica DM6000 B) using brightfield illumination and a high-speed FLIR camera (Sony Grasshopper 3; GS3-U3-32S4M-C) with a field of view of 2048 × 1536 pixels (706 μm × 530 μm). One μm diameter particles were measured with a widefield upright microscope (Leica DMI5000 M) using darkfield illumination and a high-speed FLIR camera (Sony Grasshopper 3; GS3-U3-23S6M-C) with a field of view of 1920 × 1200 pixels (1129 μm × 706 μm). Particles in the field of view were tracked in 1–3 blocks of 5 min at a framerate of 60 Hz using dedicated particle tracking software which applies two-dimensional phasor-based localization43 (link). The recorded particle trajectories were further analysed using Deep Learning based data analysis to classify unbound and bound states. Details can be found in Supplementary Notes 1 and 2 . The dose-response curves of the bound fraction, switching activity and state lifetimes were fitted with a sigmoidal curve to extract the EC50:
7
Metallographic Analysis of NiTi Instruments
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Metallographic analysis was performed on new and after SCU OCurve (n=2) and OShape (n=2) to visualise crystallographic features of the inner part of the NiTi instruments. Each instrument was sectioned and embedded at a room temperature in epoxy resin, ground and polished as described for XRD. Chemical etching was performed with a solution consisting of 60% nitric acid, 30% acetic acid and 10% hydrofluoric acid for 10 seconds in order to disclose the metallographic features (20 (link)). Optical microscope analysis (Leica DMI 5000M, Weltzlar, Germany) focused at the tip of the instruments, at 5 mm from the tip and near the handle. Micrographs between 50× and 1000× were taken.
8
Microstructural Analysis of Casting Alloys
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The metallographic samples (size: 7 × 7 × 7 mm3) were cut from the bottom of the ingots and prepared by standard grinding, polishing, and etching processes. Optical microscope (LEICA/DMI5000M, Germany) was used to analysis the microstructure of the casting alloys. A SEM (Sigma 500, Zeiss, Germany) combined with an energy-dispersive spectrometer (EDS, Oxford) was used to observe the microstructure and identify the phases present in the alloys. The 3D morphology of the Fe-rich phases was revealed by methanol and solid iodine solution used to remove the Al matrix for 3–6 h. 30 binarized optical images were used to calculate the Fe-rich phases average size. The EBSD maps and Kikuchi patterns were measured using a sample-to-detector distance of ∼ 10 mm and a voltage of 20 kV at the SEM. The sample was tilted 70° with respect to the detector. The scanning step size was set at 1 μm. The conventional procedure of counting grain boundaries in vertical and horizontal lines was used to calculate the average grain size. The samples were heated at a constant heating rate from 25 °C to 700 °C in DSC (Netzsch DSC 404, Germany) and then a constant cooling rate (10 °C/min) to 25 °C with the protection of Ar gas.
9
Immunohistochemical Analysis of Glial and Neuronal Markers
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Brain sections were rinsed with PBS, treated with 0.3% H2O2 for 15 min, and rinsed three times for 5 min each in PBS. The sections were blocked with 1% goat serum albumin for 10 min at room temperature and then incubated overnight at 4 °C with an antibody against glial fibrillary acidic protein (GFAP) (16825-1-AP, 1:1000) or microtubule-associated protein (MAP)2 (17490-1-AP, 1:100) (Proteintech). The sections were rinsed and incubated with biotin-conjugated Affinipure goat anti-rabbit IgG (H+L) (#SA00004-2, 1:500; Proteintech) for 1 h at room temperature, followed by diaminobenzidine (Solarbio; DA1010). The sections were dehydrated through an alcohol gradient, cleared in xylene, and mounted for observation with a light microscope (Leica; DMI 5000M). GFAP-positive cells were counted in each group under the same magnification and the optical density was analyzed with ImageJ software (National Institutes of Health, Bethesda, MD, USA).
10
Calcium Deposition Quantification in Bone Nodules
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To assess calcium deposition in bone nodules in the extracellular matrix, alizarin red (Beyotime, China) was used to stain the mineralized nodules in the cellular matrix. Cells were seeded in 6-well plates and treated with 0 µM and 20 µM isofraxidin for 7, 14, 21, 28, and 35 days. The medium was then removed, and the cells were washed twice with PBS, fixed in an absolute ethanol fixative for 20 min, and then removed. The fixed cells were washed twice with ionized water, and 1 mL of alizarin red was added to each well for 30 min. The solution was then rinsed twice with PBS. Mineralization results were observed under an inverted microscope (Lecia DMI5000M, Germany).
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