Fluorescence and Raman imaging was performed on the same system. A fluorescence microscope (DM2500M, Leica Microsystems Inc., Buffalo Grove, IL) equipped with an ebq 100-04 mercury lamp (Leica Microsystems Inc.), 560 nm excitation and 630 nm emission filters (Chroma 49008 ET-mCh/tR) and a 5× objective (0.12, WD=14 mm; Leica Microsystems Inc.) is integrated in the Renishaw inVia Raman imaging system (Hoffman Estates, IL), with the option to switch between the two imaging modalities. In fluorescence acquisition mode, imaging was recorded with a Lumenera Infinity 3 microscopy camera (Ottawa, Ontario, Canada). The samples were raster-scanned and stitched with Wire 3.4 software (Renishaw Inc., Hoffman Estates, IL). After fluorescence imaging, the inVia imaging system was switched to Raman acquisition mode, and SERRS images of the same samples were acquired using the StreamLine high-speed Raman imaging mode. The inVia Raman system is equipped with a 300-mW 785-nm diode laser and a 1-inch charge-coupled device detector with a spectral resolution of 1.07 cm−1 (Renishaw Inc., Hoffman Estates, IL). Raman images were generated from the raw data by applying a direct classical least square (DCLS) algorithm (Wire 3.4 software, Renishaw), which linearly fits the predefined Raman spectrum of the SERRS nanoparticle to the Raman spectra of the scanned tissues.
Dm2500m
Manufactured by Leica
Sourced in Germany, United Kingdom, Switzerland
The DM2500M is a microscope designed for routine laboratory use. It features a standard optical system with a range of magnification options. The microscope is suitable for a variety of applications requiring detailed observation and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Dfc420c, by Leica (3 mentions)
Wire 3, by Renishaw (1 mentions)
Application suite, by Leica (1 mentions)
Jsm 6400, by JEOL (1 mentions)
Model 4201, by Instron (1 mentions)
E 1030 ion sputter, by Hitachi (1 mentions)
S 5500, by Hitachi (1 mentions)
Model 5543, by Instron (1 mentions)
Microhardness tester, by Mitutoyo (1 mentions)
Supra 55, by Zeiss (1 mentions)
Mira3, by TESCAN (1 mentions)
Quantax 200, by Bruker (1 mentions)
Jsm 5900, by JEOL (1 mentions)
Jsm it300, by JEOL (1 mentions)
Raman microscope, by Renishaw (1 mentions)
Rl633, by Renishaw (1 mentions)
Sprague dawley rats, by Orient Bio (1 mentions)
Villanueva osteochrome bone staining solution, by Polysciences (1 mentions)
Nova 200, by Thermo Fisher Scientific (1 mentions)
Axis ultra dld spectrometer, by Shimadzu (1 mentions)
37 protocols using dm2500m
1
Correlative Fluorescence and Raman Imaging
2
Bone-Implant Contact Analysis of Miniscrews
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Two miniscrew-containing tibias were removed from both the 3- and 6-week groups, fixed with 10% formalin, stained in Villanueva bone, dehydrated in a series of increasing concentrations of alcohol, and embedded in methyl methacrylate. Subsequently, the blocks were cut parallel to the miniscrew axis and ground to make 40-µm thick sections. The miniscrew-bone interface was examined and bone-implant contact (BIC%) was measured on 5 threads using an optical microscope (DM 2500M, Leica Microsystems CMS GmbH) equipped with the Leica Application Suite (version 1.6.0; Leica Microsystems AG, Heerbrugg, Switzerland) under 100× magnification. BIC% values were determined as the percentage of direct contact between mineralized bone and the miniscrew surface within each thread. Five threads from two miniscrews from both groups were chosen randomly for this evaluation.
3
Failure Mode Analysis of Y-TZP and Metal Brackets
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After measuring the SBS, the bonded surface were analyzed using a light microscope (DM 2500M, Leica Microsystems, Wetzlar, Germany) at a magnification of 20× to examine the failure modes between the Y-TZP and the orthodontic metal bracket. The failure modes were then classified into adhesive failure, cohesive failure, or mixed (adhesive + cohesive) failure, and the ratios of the failure modes were calculated. Next, the failed surfaces of the Y-TZP and the metal brackets were analyzed using SEM (JSM-6400, JEOL, Tokyo, Japan).
4
Enamel-3Y-TZP Bond Strength Evaluation
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Fifty of the specimens of enamel that had been bonded to 3Y-TZP were stored in distilled water at 37 ± 1 °C for 24 h (the first test in the ISO/TS 11405 standards [17 ]; short-term storage). A universal testing machine (Model 4201, Instron, Canton, MA, USA) was then used to measure the tensile bond strength (TBS) between the 3Y-TZP and the enamel. The bonded specimens were fixed to the jig having a mini-dumbbell shape, and a tensile force was loaded at a crosshead speed of 1.0 mm/min (Figure 1 B,C).
After measuring TBS, the bonded surfaces were analyzed using a light microscope (DM 2500M, Leica Microsystems, Wetzlar, Germany) at ×20 magnification to examine the failure modes between the 3Y-TZP and the enamel. Failure modes were classified into adhesive failure, cohesive failure, or mixed failure, and the ratios of each were calculated.
After measuring TBS, the bonded surfaces were analyzed using a light microscope (DM 2500M, Leica Microsystems, Wetzlar, Germany) at ×20 magnification to examine the failure modes between the 3Y-TZP and the enamel. Failure modes were classified into adhesive failure, cohesive failure, or mixed failure, and the ratios of each were calculated.
5
Shear Bond Strength Testing of Y-TZP to Dentin
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Each specimen was mounted in the jig of a universal testing machine (Model 5543, Instron, Canton, MA, USA), and shear force was loaded onto it using a chisel-shaped metal rod at a constant crosshead speed of 0.5 mm/min until failure occurred (Figure 1 ). The SBS was calculated using the following formula:
The load was measured by recording the force at which failure occurred.
After measuring the SBS, the debonded surface was analyzed using a stereomicroscope (DM 2500M, Leica Microsystems, Wetzlar, Germany) to determine the mode of failure between the Y-TZP and bovine dentin. The failure mode was classified as adhesive, cohesive, or mixed, and the ratio of each failure mode was calculated. Representative morphologies of debonded specimens were investigated by scanning electron microscopy (SEM; S-5500, Hitachi, Tokyo, Japan) after sputtering using a gold-palladium alloy conductive layer (Ion sputter E-1030, Hitachi).
The load was measured by recording the force at which failure occurred.
After measuring the SBS, the debonded surface was analyzed using a stereomicroscope (DM 2500M, Leica Microsystems, Wetzlar, Germany) to determine the mode of failure between the Y-TZP and bovine dentin. The failure mode was classified as adhesive, cohesive, or mixed, and the ratio of each failure mode was calculated. Representative morphologies of debonded specimens were investigated by scanning electron microscopy (SEM; S-5500, Hitachi, Tokyo, Japan) after sputtering using a gold-palladium alloy conductive layer (Ion sputter E-1030, Hitachi).
6
Histopathological Evaluation of Rat Paws
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After collection of blood samples, all animals were euthanized. The right and left paws were dissected from the rats and kept intact for histological examination and the soft tissues of paw were isolated carefully. Paw tissues were fixed immediately and decalcified in ethylenediaminetetraacetic acid [21 (link)]. Then, the tissue specimens were dehydrated, embedded in paraffin, and sectioned into 5 μm portions, and stained using hematoxylin and eosin (H&E) for histopathological examination using a research microscope provided with an HD camera (Leica DM2500 M; Leica Microsystems GmbH, Wetzlar, Germany). A histopathologic score is used for inflammation, congestion, and edema on a 7-grade scale from 0 to 6 (0 = normal and 6 = the most severe inflammation).
7
Microhardness Evaluation of Dental Composites
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The Vickers hardness test was performed according to the C1327 standard using a Mitutoyo microhardness tester (Mitutoyo, Takatsu-ku, Japan) under a force of 1 kg for composite resins and 2 kg for glass ceramics for a constant indenter dwell time of 15 s. We selected two random points on each sample for hardness measurements (five samples per group). All indentation tests were performed under ambient laboratory conditions. After indentation, the hardness value of each material was obtained by calculating the diagonal length of each square indentation. Subsequently, we used an image analyzer (Leica DM 2500M, Leica Microsystems, Wetzlar, Germany) to observe the indentations.
8
Immunodetection of Renal TLR4 Expression
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Kidney samples were fixed in 10% neutral buffered formalin (4°C, overnight), embedded in paraffin, sliced into sections 5 µm thick. The paraffin sections were subsequently de-waxed in water, and incubated with 3% H2O2 for 10 min at room temperature. The sections were incubated with 3% goat serum (cat. no. 16210064; Thermo Fisher Scientific, Inc.) at room temperature for 10 min, and subsequently incubated with a primary antibody against TLR4 (1:100, cat. no. sc-13593, Santa Cruz Biotechnology, Inc.) 4°C overnight. An avidin-biotin-HRP complex immunodetection kit (Shanghai Shenggong Biology Engineering Technology Service, Ltd.) was used to detect the reaction according to the manufacturer's instructions, and the samples were examined by light microscopy at ×400 magnification (Leica DM2500M; Leica Microsystems GmbH, Wetzlar, Germany).
9
Optical Microscopy Analysis of API Crystals
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An optical microscope (Leica DM 2500M, Leica Microsystems Ltd., Heerbrugg, Switzerland) equipped with a video camera (Leica DFC420 C, Leica Microsystems Ltd., Heerbrugg, Switzerland) was used to obtain pictures of the API crystals in transmittance mode and brightfield illumination, using the 10×, 20×, and 40× Leica objective lenses.
Approximately 3 mg of the powder samples were dispersed in 3 mL of mineral oil (ACRŌS ORGANICS, Geel, Belgium) while the suspensions were analyzed as received. One or two drops of the aforementioned samples were then placed on a 76 mm × 26 mm × 1 mm microscope slide (Paul Marienfeld GmbH & Co. KG, Lauda-Königshofen, Germany), covered with a 22 mm × 22 mm cover slip (Paul Marienfeld GmbH & Co. KG, Lauda-Königshofen, Germany). Windows-based software (LAS© V4.11) was used for image acquisition and analysis.
Approximately 3 mg of the powder samples were dispersed in 3 mL of mineral oil (ACRŌS ORGANICS, Geel, Belgium) while the suspensions were analyzed as received. One or two drops of the aforementioned samples were then placed on a 76 mm × 26 mm × 1 mm microscope slide (Paul Marienfeld GmbH & Co. KG, Lauda-Königshofen, Germany), covered with a 22 mm × 22 mm cover slip (Paul Marienfeld GmbH & Co. KG, Lauda-Königshofen, Germany). Windows-based software (LAS© V4.11) was used for image acquisition and analysis.
10
Histological Assessment of Liver Injury
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The liver of each bird was harvested, stored in 10% buffered formalin, and embedded in paraffin. For histological investigations, 3 μm sections were cut, deparaffinized, dehydrated, and stained with hematoxylin and eosin (H&E) (Ma et al., 2022 (link)). The tissue sections were observed for inflammatory changes under a light microscope, Leica DM 2500 M (Leica Microsystems, Wetzlar, Germany).
Liver tissues were observed for focal inflammation, mononuclear infiltration, loss of normal architecture, and necrosis. Acute liver injury was assessed using a composite inflammation and necrosis score as described by Siegmund et al. (2002) (link). Lobular inflammation (0, no inflammation; 1, mild; 2, moderate; 3, severe), portal inflammation (0, no inflammation; 1, mild; 2, moderate; 3, severe), and necrosis (0, no necrosis; 1, ˂10% of liver parenchyma; 2, 10%–25% of liver parenchyma; 3, ˃25% of liver parenchyma) were scored and summed up to determine the overall histopathology score. Six 3 μm sections from each treatment group were scored by a trained, blinded researcher.
Liver tissues were observed for focal inflammation, mononuclear infiltration, loss of normal architecture, and necrosis. Acute liver injury was assessed using a composite inflammation and necrosis score as described by Siegmund et al. (2002) (link). Lobular inflammation (0, no inflammation; 1, mild; 2, moderate; 3, severe), portal inflammation (0, no inflammation; 1, mild; 2, moderate; 3, severe), and necrosis (0, no necrosis; 1, ˂10% of liver parenchyma; 2, 10%–25% of liver parenchyma; 3, ˃25% of liver parenchyma) were scored and summed up to determine the overall histopathology score. Six 3 μm sections from each treatment group were scored by a trained, blinded researcher.
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