Eclipse ti u

Manufactured by Nikon

Sourced in Japan, United States, France, Canada

The Eclipse Ti-U is a research-grade inverted microscope designed for advanced imaging applications. It features a high-quality optical system and a modular design to accommodate a wide range of microscopy techniques and accessories.

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613 protocols using eclipse ti u

Apoptosis Profiling of Microalgal Cells

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Briefly, the thalli on the slide were stained with 0.5% Evans blue for 10 min in the dark, and then rinsed with sterile seawater. The slides were observed under a fluorescence microscope (ECLIPSE Ti-U, Nikon, Japan). Dead cells were stained blue.
According to the manufacturer’s instructions, apoptosis was assessed using a one-step TUNEL apoptosis assay kit from Beyotime Biotechnology Company (Shanghai, China). After treatment, the slides were sealed with an antifade mounting medium and observed under a fluorescence microscope (ECLIPSE Ti-U, Nikon, Japan).

Wang Z., Lu C., Chen J., Luo Q., Yang R., Gu D., Wang T., Zhang P, & Chen H. (2022). Physiological and multi-omics responses of Neoporphyra haitanensis to dehydration-rehydration cycles. BMC Plant Biology, 22, 168.

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Visualizing HEK 293 Cell Viability

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HEK 293 cells (2 × 10⁴ cells/well), cultured in 96-well plates in DMEM media supplemented with 10% FBS, were treated with lunathrombase (2.0 μM) or medium (control) for 24 h. The cells were washed with 1x PBS, pH 7.4, stained with 5.0 μM calcein-AM (in 1x PBS, pH 7.4) and incubated for 5 min. The cells were washed with 1x PBS, pH 7.4, and visualized using an epi-fluorescence microscope at 40× magnification (Nikon ECLIPSE Ti-U, Tokyo, Japan)^{77 (link)}. For the phase-contrast images, photomicrographs were captured using a Nikon ECLIPSE Ti-U (Tokyo, Japan) camera without filter.

Gogoi D., Arora N., Kalita B., Sarma R., Islam T., Ghosh S.S., Devi R, & Mukherjee A.K. (2018). Anticoagulant mechanism, pharmacological activity, and assessment of preclinical safety of a novel fibrin(ogen)olytic serine protease from leaves of Leucas indica. Scientific Reports, 8, 6210.

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Quantifying Cell Death In Vivo and In Vitro

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For in vivo experiments, Fluoro-Jade C (FJC) staining was used to assess cell death at perihematomal region as reported previously [34 (link),35 ]. Briefly, mice were anesthesia and cardiac perfusion, and coronal brain sections were stained with FJC (AG325, Millipore). Treated brain sections were observed and photographed under a fluorescence microscope (ECLIPSE Ti–U, NIKON) at an excitation wavelength of 450–490 nm.
To assess cell death in vitro, Propidium iodide (PI) was used. Briefly, 2 × 10⁴ cells suspension was added to the 96-well plate. After the maturation, the cells were treated according to the experimental design, and the PI dye (P4170, Sigma-Aldrich) was added for 0.5 h, and the staining was observed under a microscope (ECLIPSE Ti–U, NIKON).

Xiao Z., Shen D., Lan T., Wei C., Wu W., Sun Q., Luo Z., Chen W., Zhang Y., Hu L., Zhang C., Wang Y., Lu Y., Wang P., Yang F, & Li Q. (2022). Reduction of lactoferrin aggravates neuronal ferroptosis after intracerebral hemorrhagic stroke in hyperglycemic mice. Redox Biology, 50, 102256.

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Histological Analysis of Rat Kidney

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Rats were humanely sacrificed, and their left kidneys were fixed in 10% (w/v) neutral formaldehyde, dehydrated with a graded series of alcohol, and embedded in paraffin wax. Paraffin sections (4 μm thick) of kidney were subjected to hematoxylin–eosin (HE), periodic acid-Schiff (PAS), and periodic acid-silver methenamine (PASM) staining following the standard staining protocols. The sections were imaged with a microscope (Nikon Eclipse Ti-U, Nikon Corporation, Tokyo, Japan).
Dewaxed sections were blocked with 5% bovine serum albumin (BSA) after antigen retrieval. Sections were then incubated with primary antibody against CD68 (1:100; CST, USA) followed by incubation with horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (Dako, Wuhan, China). CD68 expression was visualized by diaminobenzidine (DAB) (Dako, Wuhan, China) staining. The sections were then imaged with a microscope (Nikon Eclipse Ti-U, Nikon Corporation, Tokyo, Japan).

Hou B., He P., Ma P., Yang X., Xu C., Lam S.M., Shui G., Yang X., Zhang L., Qiang G, & Du G. (2020). Comprehensive Lipidome Profiling of the Kidney in Early-Stage Diabetic Nephropathy. Frontiers in Endocrinology, 11, 359.

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Graphene Cytotoxicity and Cell Adhesion

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Glass coverslips covered with a graphene layer (n = 20) were applied to evaluate graphene cytotoxicity to SMCs. As a control group, coverslips without graphene were used (n = 20). SMCs were seeded (20 000 cells/cm²) and cultured until confluency was reached. The first stage of analysis included evaluation of cells morphology and growth pattern (Nikon Eclipse Ti-U, Japan). Then multi-staining (Calcein/Ethidium/DAPI)(LIVE/DEAD Viability/Cytotoxicity Kit, Thermofisher, USA) was used to determine the viability of cultivated cells on the graphene covered surface. The number of viable cells was estimated as follows. The number of ethidium positive cells was subtracted from the number of DAPI positive cells. Labeled cells were counted with a confocal microscope (Nikon Eclipse Ti-U, Japonia) within the circular ROI (Region of Interest) (n = 10). The analysis was performed at 40x magnification with particle analysis function in ImageJ (https://github.com/imagej/imagej).
Glass coverslips with a graphene layer (n = 5) were applied to evaluate SMCs adherence to surface covered with graphene. As a control group, coverslips without graphene were used (n = 5). SMCs were seeded (20 000 cells/cm²). Subsequently adherent cells were counted within ROI (n = 5) 3 h, 6 h and 12 h after seeding.

Adamowicz J., Pasternak I., Kloskowski T., Gniadek M., Van Breda S.V., Buhl M., Balcerczyk D., Gagat M., Grzanka D., Strupinski W., Pokrywczynska M, & Drewa T. (2020). Development of a conductive biocomposite combining graphene and amniotic membrane for replacement of the neuronal network of tissue-engineered urinary bladder. Scientific Reports, 10, 5824.

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Isolation and Expansion of FHPCs

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The FHPCs were seeded onto type I collagen (Sigma-Aldrich)-coated 60-mm dishes at a density of 1×10⁶ viable cells. The cells were maintained in DMEM/F-12 containing 1% penicillin/streptomycin, 10 μg/ml insulin (Sigma-Aldrich), 10% fetal bovine serum (FBS), 20 ng/ml epidermal growth factor (EGF; Peprotech, Inc., Rocky Hill, NJ, USA), 10 ng/ml leukemia inhibitory factor (LIF; Sigma-Aldrich) and 2 mM L-glutamine at 37°C with 5% CO₂. The medium was changed after 24 h to remove dead and non-adherent cells, and the culture medium was subsequently changed every 1–2 days. The cells were observed daily under an inverted microscope (Nikon Eclipse Ti-U). The stellate cells were selectively detached from the dish by repeated differential digestion with TrypLE™ Express (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) at 37°C for 2.5 min. The purified FHPCs were first passaged at a ratio of 1:2 at 5–7 days after plating and passaged every 2–3 days after a generation. The passaged FHPCs were serially diluted to a density of one cell/100 μl of culture medium and replated onto uncoated 96-well plates. Colonies containing >50 cells were quantified after 2 weeks using a binocular inverted microscope (Nikon Eclipse Ti-U).

Yu L., Chen S., Luo N, & He S. (2017). The C-terminus domain of the hepatitis B virus X protein stimulates the proliferation of mouse foetal hepatic progenitor cells, although it is not required for the formation of spheroids. International Journal of Molecular Medicine, 40(2), 400-410.

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Apoptosis Induction in Cancer Cells

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HCT116 and HepG2 cells were plated at a density of 1 × 10⁵ cells/well in a 24-well plate and then incubated for 24 hours. After 24 hours, cells were treated with compound (18B) or imatinib mesylate (at IC₅₀concentration), DMSO (0.5% v/v, served as control) for 48 hours in growth medium containing 5% v/v FBS. After 48 hours, cells bright field microscopic images were captured from inverted microscope (Nikon Ti-U Eclipse, Japan). Then after, the medium was aspirated and washed with Phosphate Buffered Saline (PBS) and then cells were stained with 1 μg/mL Hoechst 33342 (HiMedia, India) [a fluorescent DNA-staining dye to detect nuclear fragmentation or chromatin condensation, features of apoptosis] for 30 minutes in dark. The morphological changes in the nucleus stained with Hoechst 33342 were captured from inverted fluorescence microscope (Nikon Ti-U Eclipse, Japan).

Neogi K., Murumkar P.R., Sharma P., Yadav P., Tewari M., Karunagaran D., Nayak P.K, & Yadav M.R. (2022). Design, synthesis and evaluation of 4,7-disubstituted 8-methoxyquinazoline derivatives as potential cytotoxic agents targeting β-catenin/TCF4 signaling pathway. Translational Oncology, 19, 101395.

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Immunocytochemistry of Neural Markers

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For immunocytochemistry, cells were cultured on glass coverslips. After washing in PBS, cells were fixed 10 min at RT with PFA 4%. For Rnd2 staining, we used the PGT-based protocol described in “Immunohistochemistry section” with a fluorescent secondary antibody. For other stainings, cells were treated with PBS – 0.01% Triton X-100 – 1% bovine serum albumin (BSA, Sigma) for 30 min and incubated overnight at 4°C with primary antibodies diluted in blocking solution: mouse anti-MAP2 (1/500, Sigma, M4403), chicken anti-nestin (1/400, Aves Labs, NES), mouse anti-Tuj1 (1/2000, Promega, G712A). Cells were then incubated with appropriate fluorescent secondary antibodies. Following this step, DAPI was added for 10 min. Images were acquired using an Eclipse Ti-U Nikon or a Leica SP5 microscope.

Kerloch T., Farrugia F., Bouit L., Maître M., Terral G., Koehl M., Mortessagne P., Heng J.I., Blanchard M., Doat H., Leste-Lasserre T., Goron A., Gonzales D., Perrais D., Guillemot F., Abrous D.N, & Pacary E. (2021). The atypical Rho GTPase Rnd2 is critical for dentate granule neuron development and anxiety-like behavior during adult but not neonatal neurogenesis. Molecular Psychiatry, 26(12), 7280-7295.

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Immunocytochemical analysis of neuronal markers

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For immunocytochemistry, cells were cultured on glass coverslips. After washing in PBS, cells were fixed 10 min at RT with PFA 4%. For Rnd2 staining, we used the PGT-based protocol described in “Immunohistochemistry section” with a fluorescent secondary antibody. For other stainings, cells were treated with PBS – 0.01% Triton X-100 – 1% bovine serum albumin (BSA, Sigma) for 30 min and incubated overnight at 4 °C with primary antibodies diluted in blocking solution: mouse anti-MAP2 (1/500, Sigma, M4403), chicken anti-nestin (1/400, Aves Labs, NES), mouse anti-Tuj1 (1/2000, Promega, G712A). Cells were then incubated with appropriate fluorescent secondary antibodies. Following this step, DAPI was added for 10 min. Images were acquired using an Eclipse Ti-U Nikon or a Leica SP5 microscope.

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oxLDL-Induced Monocyte Adhesion Assay

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EA.hy926 cells (4×10⁵ cells/ml) were seeded in six-well plates and cultured until ~90% confluence under HG conditions (25 mM) for 48 h at 37°C before being treated with oxLDL in the presence or absence of caspase-1, ROS, p38 MAPK and TXNIP inhibitors or RA at the indicated concentrations (1, 10, 50 and 100 µM) for an additional 24 h at 37°C. THP-1 cells (7×10⁵ cells per well) were labeled with the BCECF fluorescent dye for 30 min at 37°C, before being added onto the ECs and incubated for 30 min at 37°C. Following incubation, non-adherent cells were washed three times with PBS and the number of THP-1 cells adhered onto the EC cells was counted using a fluorescence microscope (Eclipse Ti-U, Nikon Corporation). Images were acquired at ×200 magnification from five randomly selected fields per well. Adhered cells were counted using ImageJ version 5.2.0 software (National Institutes of Health).

Nyandwi J.B., Ko Y.S., Jin H., Yun S.P., Park S.W., Kang K.R, & Kim H.J. (2022). Rosmarinic acid downregulates the oxLDL-induced interaction between monocytes and endothelial cells, in addition to monocyte diapedesis, under high glucose conditions. International Journal of Molecular Medicine, 49(5), 68.

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