The liver tissues were fixed with 4% paraformaldehyde (Shanghai Standard Co., Ltd., Shanghai, China) for 24 h at 40°C, and embedded in paraffin. The 5 µm slices were subsequently stained with hematoxylin-eosin and evaluated for pathological changes under a BX51 optical microscope (Olympus). Immunohistochemical staining was performed on the 4 µm paraffin-embedded sections. The slices were incubated with the previously mentioned primary antibody against FXR (1:100 dilution; Cell Signaling Technology). Following incubation with goat anti-rabbit immunoglobulin (Ig) G horseradish peroxidase-conjugated (cat. no. sc-2004; 1:1,000; Santa Cruz Biotechnology, Inc.) and goat anti-mouse IgG horseradish peroxidase-conjugated (cat. no. AP308P; 1:2,000; Sigma-Aldrich) secondary antibodies, the images were visualized using a BX51 optical microscope (Olympus). The positively stained tissue was counted using Image Pro Plus software, version 6.0 (Media Cybernetics, Silver Spring, MD, USA).
Bx51 optical microscope
Manufactured by Olympus
Sourced in Japan, United States, Germany
The BX51 is an optical microscope designed for laboratory use. It features a sturdy, ergonomic construction and provides high-quality imaging capabilities. The microscope is equipped with various optical components, including lenses and illumination systems, to facilitate detailed observation and analysis of samples.
Automatically generated - may contain errors
Lab products found in correlation
Entellan synthetic resin, by Merck Group (3 mentions)
Rm2245 rotary microtome, by Leica (2 mentions)
Dp71 digital camera, by Olympus (2 mentions)
Camedia c4040zoom camera, by Olympus (2 mentions)
Dmlp425r, by Thorlabs (2 mentions)
Uplanfl 100x 1.30 oil, by Olympus (2 mentions)
Umplanfl 100x 0, by Olympus (2 mentions)
Sld3236vf laser diode, by Thorlabs (2 mentions)
Itc4005 laser, by Thorlabs (2 mentions)
Fgl570, by Thorlabs (2 mentions)
Lumplanfl n 60x 1.00 w, by Olympus (2 mentions)
Fesh0450, by Thorlabs (2 mentions)
Rm2125rt, by Leica (2 mentions)
Rm2235, by Leica (2 mentions)
Image pro plus software version 6, by Media Cybernetics (1 mentions)
Tunel detection kit, by Beyotime (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Keygen Biotech (1 mentions)
Tritc labelled phalloidin, by Merck Group (1 mentions)
Benchmark xt automated immunohistochemistry instrument, by Roche (1 mentions)
Heto powerdry ll3000 freeze dryer, by Thermo Fisher Scientific (1 mentions)
120 protocols using bx51 optical microscope
1
FXR Expression in Liver Tissue
2
Apoptosis Detection in Paraffin Sections
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After the paraffin-embedded sections were dewaxed and dehydrated, apoptosis was detected using the TUNEL detection kit (Beyotime Institute of Biotechnology). The sections were incubated with protease K (diluted with 10 mM Tris-HCl) at 37°C for 25 min. Following 3 PBS washes and drying, the sections were added to mounting fluid and incubated at room temperature for 20 min. The sections were then washed again with PBS in triplicate and incubated with 50 µl of biotin labeling solution at 37°C for 60 min. Thereafter, the sections were washed 3 times with PBS and 2,4-diaminobutyric acid reagent was added to observe coloration under the BX-51 optical microscope (Olympus Optical Co., Ltd.). Color development was terminated by adding water, and the nuclei were stained in hematoxylin at room temperature for 1 min, dehydrated and cleared, and were then sealed with neutral gum. Each section was observed by two pathologists under a BX-51 optical microscope (Olympus Optical Co., Ltd.) and the number of TUNEL-positive cells stained brown yellow in 5 random fields was counted.
3
Characterization of Ceramic-Coated 3D Scaffolds
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Five replicas were tested per group of samples: CN, 100S, 200S, and COMPOSITE. Prior to the application of the ceramic coating, the scaffolds were morphologically characterized to ensure that there were non-significant differences between the different groups tested in terms of their main dimensions and weight. The weight of the structures was assessed by using a GR-200 analytical balance (±0.1 mg, A&D Instruments Ltd., Ahrensburg, Germany). Additionally, the height and diameter of the scaffolds, obtained as the mean value of 5 measurements per sample, were determined using an electronic calliper (±0.01 mm). After the coating process, the scaffolds were measured again following the same procedure to confirm that the pressure application and heating steps did not affect the overall dimensions of the 3D structure. The amount of ceramic additive incorporated into the scaffolds was calculated from the weight increase as a result of the coating process.
In addition, the pore size of the structures was determined, before and after coating, as the distance between strands (n = 9) by using an Olympus BX51 optical microscope (Olympus Co., Ltd., Tokyo, Japan). The thickness of the ceramic layer incorporated into the scaffolds surface was estimated as the difference between the results obtained from the pore size analysis before and after the coating process.
In addition, the pore size of the structures was determined, before and after coating, as the distance between strands (n = 9) by using an Olympus BX51 optical microscope (Olympus Co., Ltd., Tokyo, Japan). The thickness of the ceramic layer incorporated into the scaffolds surface was estimated as the difference between the results obtained from the pore size analysis before and after the coating process.
4
Visualization of RAW264.7 Cell Morphology
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RAW264.7 cells (2 × 105) were maintained on sterile glass coverslips (Corning Life Sciences Inc., Tewksbury, MA, USA) in 35 × 35 mm2 culture dishes with complete medium. After stimulation with 1 mg/mL GOS for 24 h, stimulation with 1 μg/mL LPS alone for 24 h, or pretreatment with GOS for 2 h followed by stimulation with LPS for 22 h, the cells were washed three times with PBS. Then, they were fixed with 4% paraformaldehyde for 15 min at room temperature (RT) and incubated with DAPI (KeyGEN Biotech, Nanjing, China) for 10 min at RT or with TRITC-labelled phalloidin (1:200) (Sigma-Aldrich, St. Louis, MO, USA) in the dark for 20 min at 4 °C for staining of the F-actin fibers. For examination of cell morphology and the actin cytoskeleton observation, fluorescence and dark-field microscopy were performed using an upright Olympus BX51 optical microscope (Olympus Corporation, Tokyo, Japan), and fluorescence and differential interference contrast microscopy were performed using an Olympus FV1000 confocal scanning laser microscope (Olympus Corporation, Tokyo, Japan).
5
Histological Evaluation of Liver Steatosis
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Formalin-fixed and paraffin-embedded liver sections were stained with hematoxylin and
eosin (HE) and Masson's trichrome for histopathological examination. The severity of
steatosis was graded by the extent of parenchymal involvement. Grade 0, <5% of the
parenchyma was involved; grade 1, 5 to 33%; grade 2, 34 to 66%; grade 3, >66%.
Other parameters assessed were the presence or absence of inflammation and fibrosis
score (17 (link)).
The histological slides were analyzed and photographed with an Olympus¯ BX
51 optical microscope (Olympus Corporation, Japan). The pathologist who evaluated the
sections and performed the histological assessments was unaware of the treatment
groups.
eosin (HE) and Masson's trichrome for histopathological examination. The severity of
steatosis was graded by the extent of parenchymal involvement. Grade 0, <5% of the
parenchyma was involved; grade 1, 5 to 33%; grade 2, 34 to 66%; grade 3, >66%.
Other parameters assessed were the presence or absence of inflammation and fibrosis
score (17 (link)).
The histological slides were analyzed and photographed with an Olympus¯ BX
51 optical microscope (Olympus Corporation, Japan). The pathologist who evaluated the
sections and performed the histological assessments was unaware of the treatment
groups.
6
Immunohistochemical Analysis of Tumor Samples
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Retrieved tumors and organs from treated mice were immersed in 4% buffered formaldehyde, and 24 h later washed, dehydrated, and embedded in paraffin. Hematoxylin–eosin (H&E) staining was performed on 4 μm sections mounted on polylysine slides. Tissues sections were observed under the OLYMPUS BX51optical microscope (Olympus Corporation, Tokyo, Japan). For IHC staining, tumor and organs slices (2 μm size) were deparaffinized and pre-treated with the Epitope Retrieval Solution 2 (EDTA-buffer, pH 8.8) at 98 °C for 20 min. After washing steps, peroxidase blocking was carried out for 10 min using the Bond Polymer. All procedures were performed using the Benchmark XT-Automated Immunohistochemistry instrument (Ventana Medical Systems, Oro Valley, AZ, USA). Tissues were again washed and then incubated with the primary antibody directed against Ki67 (Dako, clone: MIB-1; 1:150). Subsequently, tissues were incubated with polymer for 10 min and developed with DAB-Chromogen for 10 min. The experiments were repeated at least three times.
7
Deer Antler Homogenization and Preparation
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The deer antler used in the following experiments was the same as the ones prepared as previously described [22 (link)]. Antler tissue was collected from three anesthetized 4-year-old Chinese sika deer at the Shuangyang Deer Farm in Changchun, China. Deer antlers were fully rinsed with cold MilliQ water and thoroughly homogenized using a high-speed tissue homogenizer (Voshin, China). The homogenate was centrifuged at 12,000 × g and 4°C for 30 min. The supernatant was filtered through a 0.45 μm Hollow Fiber Cartridge (GE Healthcare, Chicago, IL). The filtrate was freeze-dried by a Heto PowerDry LL3000 Freeze Dryer (Thermo, Waltham, MA) and stored at −80°C. When the mice need to be administered by gavage, the velvet antler was dissolved in pure water. FA1004N electronic balance (Shanghai Precision Scientific Instrument Co., Ltd.), Leica RM2255 paraffin microtome (Leica, Germany), Leica EG1140 paraffin-embedding machine (Leica, Germany), Olympus BX51 optical microscope (Olympus, Japan), and NIS-ELEMNT BR image analysis system (Japan NIKON Company) were used.
8
Leaf Anatomy Quantification
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After four months of light control, five plants were selected from each treatment, and then selected from each plant one mature leaf with the same light direction, light time and growth trend. A double-sided blade was used to cut a 5 mm× 5 mm leaf along the main vein at the base 1/3 of the leaf, fixed using a Formalin-Aceto-Alcohol fixative on site. The slides were prepared using the paraffin section method, ethanol, xylene gradient dehydration, wax immersion, and embedding. The section thickness was 6-8 µm, and safranine-fast green double stain was used along with a neutral gum-sealing piece. The slices were observed and photographed using the Olympus-Bx 51 optical microscope (Olympus Corporation, Tokyo, Japan). The thickness of the upper epidermis, lower epidermis, palisade tissue, spongy tissue, and leaf were measured using Image-Proplus 6.0 (Media Cybernetics, Rockville, MD, USA), and the ratio of palisade tissue thickness to spongy tissue thickness was calculated. Each treatment was selected using a 15-field observation, and the average was calculated [34] .
9
Histological Examination of Hepatocellular Carcinoma
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Macroscopically visible HCC nodules greater than one mm in diameter on the surface of each liver were recorded. Parts of the collected livers were fixed in 10% v/v neutral buffered formalin for 24 h and embedded in paraffin. Histological changes were examined in 5-μm-thick sections stained with haematoxylin and eosin (H&E). HCC was graded as described by Bosman, et al.35 . Images were taken using a BX51 Olympus optical microscope (Olympus Corporation, Tokyo, Japan).
10
Kidney Histopathological Assessment Protocol
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Tubular injury was scored in kidney sections (4 μm) stained with hematoxylin–eosin using the modified Kinomura et al. scoring system [39 (link)]. Briefly, twenty random fields were examined with a light microscope (BX51 Olympus optical microscope, Olympus cooperation, Tokyo, Japan) at a magnification of X400 and quantified for histopathological changes as swelling, desquamation from the tubular basement membrane, necrosis, and vacuolar degeneration. Scores from 0 to 5 were assigned according to the percentage of the tubular histopathological changes (0: normal; 1: <20% 2: 20–40%; 3: 40–60%; 4: 60–80%; and 5: 80–100%).
Interstitial fibrosis was quantified in sections stained with Masson’s trichrome using a color image analyzer (J 1.32) avoiding blood vessels [40 (link)].
Interstitial fibrosis was quantified in sections stained with Masson’s trichrome using a color image analyzer (J 1.32) avoiding blood vessels [40 (link)].
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