Antibodies for immunohistochemical analysis were as follows: mouse monoclonal anti-CA II (sc-48351; Santa Cruz, Barcelona, Spain) at a dilution of 1:50 and rabbit polyclonal anti-CHOP (GADD 153 (F168): sc575; Santa Cruz, Barcelona, Spain) at a dilution of 1:100. Liver samples were fixed and embedded in paraffin. Antigen retrieval was performed by incubating samples with 10 mM sodium citrate. After blocking, the tissue sections were incubated overnight with the antibody. Sections were then incubated with biotinylated goat anti-mouse IgG (1:200) for CA II or with biotinylated goat anti rabbit IgG (1:200) for CHOP followed by conjugated horseradish peroxidase-streptavidin (VECTASTAIN Elite ABC Reagent). Slides were counterstained with hematoxylin and mounted with DPX (Sigma, Madrid, Spain). Images were taken with Nikon Eclipse E1000 microscope (Nikon, Lewisville, TX) using Cell B imaging software (Olympus America, Center Valley, PA; USA).
Eclipse e1000 microscope
Manufactured by Nikon
Sourced in Japan, United States, Germany
The Nikon Eclipse E1000 is a microscope designed for laboratory use. It features a modular design that allows for customization to meet specific research needs. The Eclipse E1000 provides high-quality imaging capabilities, but a detailed description of its core function is not available while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Digital sight camera, by Nikon (3 mentions)
Hematoxylin and eosin, by Muto Pure Chemicals (3 mentions)
Alexa 568 red, by Thermo Fisher Scientific (2 mentions)
Nis elements software, by Nikon (2 mentions)
Evolution qei monochrome digital camera, by Media Cybernetics (2 mentions)
Primary antibody against microtubule associated lc3, by MBL Life Science (2 mentions)
β catenin, by BD (2 mentions)
Oct optimum cutting temperature compound, by Sakura Finetek (2 mentions)
Alexafluor conjugated anti igg antibodies, by Thermo Fisher Scientific (2 mentions)
Uea 1, by Vector Laboratories (2 mentions)
Clone rm4 5, by Thermo Fisher Scientific (2 mentions)
Clone 53 6, by Thermo Fisher Scientific (2 mentions)
Tcs sp8, by Leica (2 mentions)
Eclipse ti2, by Nikon (2 mentions)
Rat anti e cadherin, by Thermo Fisher Scientific (2 mentions)
Cell b imaging software, by Olympus (1 mentions)
Dibutylphthalate polystyrene xylene (dpx), by Merck Group (1 mentions)
Anti caspase 1, by Thermo Fisher Scientific (1 mentions)
Anti nlrp3, by Thermo Fisher Scientific (1 mentions)
Anti lc3, by MBL Life Science (1 mentions)
46 protocols using eclipse e1000 microscope
1
Immunohistochemical Analysis of Liver Samples
2
Cardiomyocyte Autophagy Evaluation
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To observe autophagic activity in the cardiomyocytes, sections immunostained with primary antibodies at 10 μg/mL were anti‐LC3 (MBL International, Woburn, MA, US), anti‐NLRP3 (Thermo Fisher Scientific, Pleasanton, CA, US), anti‐Caspase‐1 (Thermo Fisher Scientific, Pleasanton, CA, US), and followed by Alexa 568 (red, Molecular Probes, Sunnyvale, CA, US). Nuclei were stained with 4′‐6‐diamidino‐2‐phenylindole. All immunostained sections were assessed under a Nikon Eclipse E1000 microscope and Nikon Digital Sight Camera (Nikon Instruments, Tokyo, Japan).
3
Visualizing Fungal Spore Morphology
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MM agar slides were prepared by pipetting 1 ml agar containing MM. MM agar slides were inoculated with spores and grown at 30 °C in petri dishes until analysis. Live cell images were captured with a cooled Evolution QEi monochrome digital camera (Media Cybernetics Inc.) mounted on a Nikon Eclipse E1000 microscope (Nikon). Images were captured using a Plan-Fluor ×100, 1.30 numerical aperture objective lens. The illumination source was a 103-watt mercury arc lamp (Osram). The fluorophore mRFP was visualised using a band pass RFP filter (EX545/30, EM620/60 combination filter; Nikon). Each slide was scanned manually, and representative images were captured to document the morphological phenotype and fluorescence pattern of each strain. Red colour was added to each image where a fluorescence signal was obtained using image processing in ImageJ.
4
Quantitative Assessment of Autophagy and Apoptosis in Cardiomyocytes
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After paraffin removal, the sections were incubated with a primary antibody against microtubule-associated LC3 (MBL International, Woburn, MA). To observe the autophagic activity in cardiomyocytes, sections immunostained with anti-LC3 followed by Alexa 568 (red; Molecular Probes) were also labeled with anti-myoglobin antibody (DAKO Japan, Kyoto, Japan) followed by Alexa 488 (green; Molecular Probes, Sunnyvale, CA). These sections were then counterstained with Hoechst 33342 and observed under Nikon Eclipse E1000 microscope and Nikon Digital Sight Camera (Nikon Instruments, Tokyo, Japan).
For in situ terminal deoxyuridine triphosphate nick end-labeling (TUNEL), tissue sections were stained with Fluorescein-FragEL (Oncogene Research Products, Boston, MA). Quantitative assessments were performed in 50 randomly chosen high-power fields (600) using an NIS-Element imaging software (Nikon Instruments, Tokyo, Japan).
For in situ terminal deoxyuridine triphosphate nick end-labeling (TUNEL), tissue sections were stained with Fluorescein-FragEL (Oncogene Research Products, Boston, MA). Quantitative assessments were performed in 50 randomly chosen high-power fields (600) using an NIS-Element imaging software (Nikon Instruments, Tokyo, Japan).
5
Hematolymphoid Tissue Imaging and Analysis
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Images of normal human hematolymphoid tissue and TMA immunohistochemical staining results were acquired using a Nikon Eclipse E1000 microscope (Nikon, Tokyo, Japan) equipped with ×4, ×10, ×20, and ×40, and ×60 objective lenses with numerical apertures ranging from 0.05 to 0.90. Images were captured with a SPOT flex mosaic 15.2 digital camera and software (Diagnostic Instruments, Sterling Heights, MI, USA). Immunofluorescence images were acquired using a Nikon Eclipse E800 microscope and a DXM1200C Nikon digital camera (Nikon, Tokyo, Japan). Digitized images were processed using Adobe Photoshop software (Adobe Systems, San Jose, CA, USA).
6
Quantifying Muscle Capillary Density
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Muscle cryosections were air dried and incubated in blocking solution (1% BSA in PBS) at room temperature for 2 h. Samples were then incubated with isolectin conjugated with Alexa Fluor 568 (capillaries, 1:500, Life Technologies, I21412) and wheat germ agglutinin conjugated with Alexa Fluor 350 (sarcolemma, 1:100, Life Technologies, W11263) in PBLEC for 1 h. Capillary density was quantified by manually counting of isolectin-positive signals on 5-6 images from each cryosection (20x) with a Nikon Eclipse E1000 microscope (Nikon, Boerhavedorp, Germany) using the NIS Elements software (Nikon Instruments Europe BV, Amsterdam, The Netherlands). Analyses were performed by investigators who were blinded to the experimental groups.
7
Extraction and Identification of Soil Nematodes
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Each soil sample was homogenized in 90 ml of 50 mM sodium phosphate buffer (pH 7.0) by agitation at 200 rpm for 1 h. To recover the nematodes, each soil suspension was placed on four layers of tissue paper in a Baermann funnel filled with distilled water. Extracts (about 2 ml) were collected every 2 days, over a 6-day period. A 1-mL aliquot was placed on a small Syracuse dish, and total nematodes were counted for each sample using a dissecting microscope at 32× magnification. Then, the nematodes were placed on slide glasses, killed by heat, and fixed by the addition of double-strength formaldehyde-acetic acid solution (100 ml 40% formaldehyde; 10 ml glacial acetic acid; and 390 ml distilled water). Nematode specimens were viewed with a Nikon Eclipse E1000 microscope (Nikon Corp., USA) from 400× to 1,000× magnification. Finally, the specimens were separated into two trophic groups, fungivores and others, by the presence of a stylet.
8
Foam Cell Quantification Protocol
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Cells were fixed in 4% buffered formalin for 10 min, washed in distilled water, rinsed in 60% isopropanol, and stained with 0.2% Oil Red O for 15 min. Stained cells were then cleaned in 60% isopropanol and were counterstained with haematoxylin. The aqueous mounting agent Aquatex (Merck, Summit, NJ, USA) was used to mount cells for light microscopic observations. For fluorescence analysis, cells were mounted in 10% glycerol in 20 mM Tris-HCl (pH 7.4) with 4′,6-diamidino-2-phenylindole (DAPI). Foam cells were observed via light or fluorescence microscopy (excitation wavelength 510–560; emission wavelength 575–590) at × 400 magnification with a Nikon Eclipse E1000 microscope (Nikon, Tokyo, Japan). Cells with more than 10 lipid droplets were defined as foam cells and the percentage of foam cells formed in each condition was determined according to a previously described method67 (link). The lipid content of macrophages was also determined by means of a colorimetric method. Details of the procedure can be found in the Supplementary Information online.
9
Quantifying Succinate Dehydrogenase Activity
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Muscle cryosections were air dried and incubated with 25% V/V Tris-HCl (50mM, pH7.4), 25% V/V Na-succinate (0.2M), 5% V/V MgCl2*6H2O (50mM) and 45% V/V nitro blue tetrazolium chloride (1.223mM) for 1 h at 37 °C in a humid chamber. Images from the whole cross-section (10x) of n=6/condition were acquired with Nikon Eclipse E1000 microscope (Nikon, Boerhavedorp, Germany) using the NIS Elements software (Nikon Instruments Europe BV, Amsterdam, The Netherlands). The entire cryosection was reconstituted using Microsoft Image Composite Editor (Microsoft, Cambridge, USA). The blue-purple color intensity was quantified to assess the succinate dehydrogenase activity in at least n=50 oxidative fibers per cryosection using the ImageJ software (National Institutes of Health, Bethesda, MD, USA).
10
Quantitative Immunohistochemistry of Cardiac Fibroblasts
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At the end of the 72-hour incubation period with the indicated treatments, parallel cultures of human cardiac fibroblasts were fixed in ice-cold methanol at Ϫ20°C (for elastin), and in paraformaldehyde at room temperature (for collagen) for 30 minutes, followed by a PBS wash. Next, the cultures were then incubated with 10-g/mL polyclonal antibody to tropoelastin or 10-g/mL polyclonal antibody to collagen type I for 1 hour. All cultures were washed and then incubated for an additional hour with fluorescein-conjugated goat antirabbit or fluorescein-conjugated rabbit antigoat secondary antibodies to detect elastin. Parallel cultures incubated with secondary antibody alone were used as a negative control. Nuclei were counterstained with red-fluorescent propidium iodide. All of the immune-stained cultures were mounted in Elvanol and examined with a Nikon Eclipse E1000 microscope attached to a cooled charge-coupled device camera (Nikon Digital Sight DS-Qi1MC; Nikon Corp) and NIS-Element imaging analysis system (Nikon Instruments, Inc Canada). The collected pictures were then analyzed with the computer-generated morphometric analysis system, in which the Image-Pro Plus software (Media Cybernetics) estimates the proportion of areas marked with green fluorescence, in relation to the entire (1 mm 2 ) analyzed field, as previously described (23, 26) .
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