Eclipse te2000 u

Manufactured by Nikon

Sourced in Japan, United States, United Kingdom, Germany, Italy, Canada

The Nikon Eclipse TE2000-U is a versatile inverted microscope designed for a wide range of applications in research and laboratory settings. It features advanced optics and a modular design to accommodate various accessories and imaging techniques. The core function of the Eclipse TE2000-U is to provide high-quality, detailed observations of specimens under different illumination and contrast methods.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (18 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (10 mentions) Triton x 100, by Merck Group (9 mentions) Hoechst 33342, by Thermo Fisher Scientific (8 mentions) Orca er, by Hamamatsu Photonics (6 mentions) Hoechst 33342, by Merck Group (6 mentions) Nis element, by Nikon (6 mentions) Plan fluor, by Nikon (5 mentions) Matrigel, by BD (5 mentions) Lambda ls system, by Sutter Instruments (5 mentions) Nis elements br 3, by Nikon (5 mentions) Ds fi2, by Nikon (4 mentions) Digital sight ds fi1, by Nikon (4 mentions) Syringe pump, by Harvard Apparatus (4 mentions) Super fluor 20 0.75 na, by Teledyne (4 mentions) Metafluor 6, by Molecular Devices (4 mentions) Lambda 10 2 optical filter change, by Sutter Instruments (4 mentions) Bovine serum albumin (bsa), by Merck Group (4 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions) Alexa fluor 488, by Thermo Fisher Scientific (4 mentions)

691 protocols using eclipse te2000 u

Histopathological Evaluation of Kidney Injury

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Kidneys were removed, fixed in 4% formalin for 24 hours, and embedded in paraffin. Sections of 3 μm thick tissues were cut and stained with hematoxylin & eosin. Morphological changes in the kidney were evaluated using a light microscope (Nikon Eclipse TE2000-U, Japan) at ×200 magnification. The degree of foaming and the detachment of renal tubular cells were scored on a scale of 0–4 as follows: no injury (0); mild: 0%–25% (1); moderate: 25%–50% (2); severe: 50%–75% (3); and very severe: 75%–100% (4).^{16 (link)} For immunohistochemical staining, the sections were incubated with primary rabbit polyclonal antibodies, including anti-HIF-α (bs-20399R; Bioss Biological, Ltd, Beijing, China), anti-HE4 (bs-4626R; Bioss Biological, Ltd), and anti-phospho-NF-κB (p65) (bs-3485R; Bioss Biological, Ltd), at a 1:100 dilution and incubated at 4°C overnight. The corresponding secondary antibodies (biotinylated goat anti-rabbit antibody from Bioss Biological, Ltd) were added and incubated at 37°C for 30 minutes, and diaminobenzidine was added for color development. Negative controls were prepared by omitting the primary antibody. After the sections were counterstained with hematoxylin and dehydrated, we observed the specimens using a light microscope (Nikon Eclipse TE2000-U, Nikon, Japan), and a brown–yellow color in the field of view represented a positive reaction.^{17 (link)}

Huang X., Guo X., Yan G., Zhang Y., Yao Y., Qiao Y., Wang D., Chen G., Zhang W., Tang C, & Cao F. (2022). Dapagliflozin Attenuates Contrast-induced Acute Kidney Injury by Regulating the HIF-1α/HE4/NF-κB Pathway. Journal of Cardiovascular Pharmacology, 79(6), 904-913.

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Cell Viability and Cell Death Assays

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Cell viability was evaluated using the metabolic dye 3-(4,5-dimethlthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT). The assays were carried out with 4.5 × 10 4 vCTB cells in each well of 96-well plates treated or not with CPF during 61 h. Results are expressed as percentage of cell viability relative to 0.04% v/v DMSO (control).
Acridine orange (AO/N,N,N′,N′-tetramethylacridine-3,6-diamine) and ethidium bromide (EB) staining were performed in vCTBs exposed to CPF during 19 or 61 h. After treatments, cells were washed with phosphate buffered saline (PBS) and 20 μL of a 1/10 dilution (in PBS) of AO stock (100 μg/mL) were added and cells were incubated during 5 min. After washing, cells were observed and images registered with an epifluorescent microscope (Nikon Eclipse TE2000-U). After, 20 μL of a 1/ 10 dilution of EB stock (100 μg/mL) in PBS were added and cells were incubated during 5 min. Washed cells were observed and registered with an epifluorescent microscope (Nikon Eclipse TE2000-U). The assays were done as quickly as possible to prevent cell death from causes beyond the specific treatments. Positive staining controls were performed to discriminate apoptotic and necrotic cells (data not shown).

Ridano M.E., Racca A.C., Flores-Martin J.B., Fretes R., Bandeira C.L., Reyna L., Bevilacqua E., Genti-Raimondi S, & Panzetta-Dutari G.M. (2017). Impact of chlorpyrifos on human villous trophoblasts and chorionic villi. Toxicology and applied pharmacology, 329.

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Quantification of ROS and Cell Death in HLECs

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ROS production and cell death were quantified by the methods described previously (Elanchezhian et al., 2012a (link); Palsamy et al., 2012 (link)). Briefly, the HLECs treated with 20 mM VPA was stained for ROS production by 2′,7′-dichlorodihydrofluorescein diacetate (H₂-DCFH-AD) (Invitrogen) for 30 min at 20°C. The cells were washed twice with PBS and subjected to fluorescent microscopic imaging (Nikon, Eclipse TE2000-U) with a green filter. The cell death in HLECs treated with 20 mM VPA was also determined by using viability/cytotoxicity assay kits (Biotium Inc.) for 30 min, and were washed twice with PBS and subjected to fluorescent microscopic imaging (Nikon, Eclipse TE2000-U) with a red filter.

Palsamy P., Bidasee K.R, & Shinohara T. (2014). Valproic acid suppresses Nrf2/Keap1 dependent antioxidant protection through induction of endoplasmic reticulum stress and Keap1 promoter DNA demethylation in human lens epithelial cells. Experimental eye research, 121, 26-34.

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Assessing Nrf2-Dependent Viability and ROS in Diabetic LECs

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LECs isolated from diabetic Nrf2^-/- and Nrf2^+/+ C57BL/6 mice as well as HLECs were treated with MGO and were stained for cell viability and cell death using viability/cytotoxicity assay kits (Biotium Inc., Hayward, CA) for 30 min. After washing twice with PBS, they subjected to fluorescent microscopic imaging (Nikon, Eclipse TE2000-U) with a green filter (450-490 nm) for viable cells and with a red filter (510-560 nm) for dead cells, respectively. This viability/cytotoxicity assay kit is consisted of calcein AM and ethidium homodimer-III (EthD). Calcein AM is a non-fluorescent substrate for cell-permeate esterase and is cleaved by intracellular esterases to form highly fluorescent calcein, which is retained in the live cells and imparts intense green color fluorescence. EthD, a membrane-impermeable fluorescent dye, undergoes a fluorescence enhancement upon binding with nucleic acids and imparts red color fluorescence. EthD is only able to enter into the dead cells, because the plasma membranes of those cells are compromised. The cytosolic ROS level in LECs treated with MGO was also determined by 2′,7′-dichlorodihydrofluorescein diacetate (H₂-DCFH-AD) (Invitrogen, Carlsbad, CA) for 30 min at 20°C. The cells were washed twice with PBS and subjected to fluorescent microscopic imaging (Nikon, Eclipse TE2000-U, Tokyo, Japan) with a green filter (450-490 nm) [45 (link), 46 (link)].

Palsamy P., Bidasee K.R., Ayaki M., Augusteyn R.C., Chan J.Y, & Shinohara T. (2014). Methylglyoxal induces endoplasmic reticulum stress and DNA demethylation in the Keap1 promoter of human lens epithelial cells and age-related cataracts. Free radical biology & medicine, 72, 134-148.

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3T3-L1 and RAW264.7 Cell Differentiation

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The seeding densities (1 × 10⁶ cells/mL) of 3T3-L1 preadipocytes and RAW264.7 macrophages (1 × 10⁴ cells/mL) were adjusted using a disposable haemocytometer-based cell counter (SKC Co. Ltd., Seoul, Korea) with the aid of an inverted microscope (Eclipse TE2000-U, Nikon, Tokyo, Japan). Cells/hydrogel mixtures were plotted using the cell plotting system. The cells in the plotted hydrogel were observed by a live/dead cell assay kit (Invitrogen, Carlsbad, CA) using fluorescence microscopy (Eclipse TE2000-U, Nikon, Tokyo, Japan). Fabricated scaffolds were differentiated using the differentiation medium containing 20 μg/mL insulin, 0.5 mM isobutylmethylxanthine and 1 μM dexamethasone for 3 days, which was then replaced with a medium containing 20 μg/mL insulin for 2 days. Cells were fully differentiated, and the medium was exchanged with an insulin-free medium containing 10% FBS.

Lee S.Y., Park S.B., Kim Y.E., Yoo H.M., Hong J., Choi K.J., Kim K.Y, & Kang D. (2019). iTRAQ-Based Quantitative Proteomic Comparison of 2D and 3D Adipocyte Cell Models Co-cultured with Macrophages Using Online 2D-nanoLC-ESI-MS/MS. Scientific Reports, 9, 16746.

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Microgel Scaffold Characterization of MSCs

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Small and large microgel scaffolds composed of 6% PEG-VS were seeded with MSCs at 5 million cells mL⁻¹. Images were acquired at 12, 24, and 48 hr with brightfield microscopy (Nikon Eclipse TE2000U). Aggregates from large microgels were collected by pipetting the scaffolds up and down, followed by pipetting the solution through a 100 μm sieve. Aggregates were washed with PBS, stained with a live/dead assay per the manufacturer’s protocol (Thermo Fisher), and fluorescent images taken using the Nikon Eclipse TE2000U. For confocal microscopy images, cells were stained and imaged with DAPI and phalloidin as described above. Aggregate size was measured in ImageJ using the line and measurement tools.

Bohlson S.S., Strasser J.A., Bower J.J, & Schorey J.S. (2001). Role of Complement in Mycobacterium avium Pathogenesis: In Vivo and In Vitro Analyses of the Host Response to Infection in the Absence of Complement Component C3. Infection and Immunity, 69(12), 7729-7735.

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Evaluating FITC-FA-DEX-VBL-SPION Cell Uptake

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FA-DEX-VBL-SPION was functionalized with fluorescein 5(6)-isothiocyanate (FITC) to evaluate its cell internalization efficiency using a fluorescence microscope (Nikon Eclipse TE2000-U). The Fluorescein Isothiocyanate (FITC)-dextran was synthesized by coupling fluoresceinyl isothiocyanate (5-isomer) to dextran polymer. FITC was conjugated randomly to hydroxyl groups of dextran at a frequency of 0.003 to 0.02 moles of FITC per mole of glucose. The cells were treated with 5 μg of FITC-FA-DEX-VBL-SPION for 3 h. Subsequently, the nanocomposite-containing medium was discarded, and the cells were washed with PBS. Photomicrographs were acquired using a fluorescence microscope (Nikon Eclipse TE2000-U).

Albukhaty S., Al-Musawi S., Abdul Mahdi S., Sulaiman G.M., Alwahibi M.S., Dewir Y.H., Soliman D.A, & Rizwana H. (2020). Investigation of Dextran-Coated Superparamagnetic Nanoparticles for Targeted Vinblastine Controlled Release, Delivery, Apoptosis Induction, and Gene Expression in Pancreatic Cancer Cells. Molecules, 25(20), 4721.

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Liver Fibrosis Quantification Protocol

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Liver tissues were fixed in 10% formalin and embedded in paraffin. Five-micron- thick sections were stained with haematoxylin-eosin (H&E), Masson and Sirius red stains. Images were acquired by light microscopy (Nikon Eclipse TE2000-U, Nikon, Japan), and the degree of liver fibrosis was quantified using Image-Pro Plus 6.0 software. For immunohistochemical staining, the paraffin sections were incubated with an anti-TGF-β1 and p-Smad2 antibodies and a biotinylated secondary antibody, followed by incubation with an avidin-biotin-peroxidase complex. Then, the signals were visualised using DAB. Images were acquired by light microscopy (Nikon Eclipse TE2000-U, Nikon, Japan).

Liu M., Xu Y., Han X., Yin L., Xu L., Qi Y., Zhao Y., Liu K, & Peng J. (2015). Dioscin alleviates alcoholic liver fibrosis by attenuating hepatic stellate cell activation via the TLR4/MyD88/NF-κB signaling pathway. Scientific Reports, 5, 18038.

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Multi-Scale Histological Imaging Protocol

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Low magnification Images of Masson’s trichrome stained samples were collected using the 10x objective on a Nikon Eclipse TE2000-U with a Nikon Digital Sight DS-2Mv Digital Camera (Nikon Inc., Melville, New York). A 6 X 6 array of higher magnification images was taken by stitching individual 20x fields together. Images of CD31 and CD68 stained samples were collected with the 20x objective on a Nikon Eclipse TE2000-U with a Nikon Digital Sight DS-2Mv Digital Camera (Nikon Inc., Melville, New York). Images were taken by stitching individual 20x images to generate 6x10 large-image fields. Illumination at 488 nm was used to image CD68 and CD31 receptors by capturing the 525 nm emission from the counterstained cell nuclei (Hoechst Dye 33342 stain).

Vallejo-Heligon S.G., Brown N.L., Reichert W.M, & Klitzman B. (2015). Porous, Dexamethasone-Loaded Polyurethane Coatings Extend Performance Window of Implantable Glucose Sensors in vivo. Acta biomaterialia, 30, 106-115.

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Cytotoxicity and Autophagy Induction by LCA

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PC-3 and DU-145 cells were added to 24-well plates at a density of 1 × 10⁵ cells/well in 1 ml of complete medium. After 24 h, cells were treated with several concentrations of LCA (0, 1, 3, 10 and 30 µM) in fresh medium containing 2% stripped FBS and another 24 h later, Hoechst 33342 (Sigma-Aldrich) and propidium iodide (Invitrogen, Carlsbad, CA, USA) were each added at a concentration of 1 µg/ml per well. After a 15 min incubation at 37 °C, cells were observed and counted under a Nikon Eclipse (TE-2000U) inverted fluorescence microscope at 20× magnification. Hoechst- and propidium iodide-positive cells were made visible using filter cubes with excitation wavelengths of 330–380 nm and 532–587 nm, respectively. To measure autophagy, PC-3 cells were exposed to LCA (0, 3, 10, 30 and 50 µM) for 24 h and then stained with Hoechst 33342 and 2 µL of Cyto-ID^® Green Detection Reagent (ENZ-51031-K200; Enzo Life Science, Farmingdale, NY, USA). After a 15 min incubation at 37 °C, cells were observed and counted under a Nikon Eclipse (TE-2000U) inverted fluorescence microscope at 20× magnification.

Gafar A.A., Draz H.M., Goldberg A.A., Bashandy M.A., Bakry S., Khalifa M.A., AbuShair W., Titorenko V.I, & Sanderson J.T. (2016). Lithocholic acid induces endoplasmic reticulum stress, autophagy and mitochondrial dysfunction in human prostate cancer cells. PeerJ, 4, e2445.

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