MCF-7 cells (5 × 104 cells/well) were seeded in 24 well plates, overnight. After incubation, the cells were then exposed to various concentrations of As2O3 (0, 4, 11, 16 and 32 µM), cobalt chloride (50 and 100 µM) and curcumin (50 and 100 µM) for 24 h. After treatment, the cells were washed with sterile 1 × PBS and fixed with 3.7% paraformaldehyde for 10 min at room temperature. The cells were then washed twice with sterile 1 × PBS and stained with DAPI (5 µg/mL) for 15 min in the dark, at room temperature. Following incubation, the cells were washed twice with sterile 1 × PBS. Morphological changes of MCF-7 cells in each group were observed under the Olympus CKX53 Inverted Microscope, (Olympus, Shinjuku, Tokyo, Japan) and Eclipse Ti-U fluorescence microscope (Nikon Instruments Inc., Melville, NY, USA), for light and fluorescence microscopy, respectively. The images were captured with DSRI-1 camera and an LC-30 camera set, respectively.
Eclipse ti u fluorescence microscope
Manufactured by Nikon
Sourced in Japan, United States
The Eclipse Ti-U fluorescence microscope is a high-performance instrument designed for advanced imaging applications. It features a modular and flexible design, allowing for the integration of various imaging techniques and components. The Eclipse Ti-U is capable of capturing detailed images of fluorescently labeled samples, enabling researchers to study cellular and molecular processes in a wide range of scientific disciplines.
Automatically generated - may contain errors
Lab products found in correlation
Spot rt digital camera, by Nikon (6 mentions)
Hoechst 33342, by Thermo Fisher Scientific (6 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (4 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (4 mentions)
Calcein am, by Thermo Fisher Scientific (3 mentions)
Triton x 100, by Merck Group (2 mentions)
Ds ri1 camera, by Nikon (2 mentions)
Fluorescence mounting medium, by Agilent Technologies (2 mentions)
Ckx53 inverted microscope, by Olympus (1 mentions)
Ab108422, by Abcam (1 mentions)
Donkey anti rabbit igg, by Santa Cruz Biotechnology (1 mentions)
Goat anti rabbit igg555, by Abcam (1 mentions)
Ab192890, by Abcam (1 mentions)
Ab7817, by Abcam (1 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Sybr green 1 nucleic acid gel stain, by Thermo Fisher Scientific (1 mentions)
Nis elements basic research software, by Nikon (1 mentions)
A2547, by Merck Group (1 mentions)
Prolong gold antifade with dapi, by Thermo Fisher Scientific (1 mentions)
α sma, by Merck Group (1 mentions)
58 protocols using eclipse ti u fluorescence microscope
1
Cellular Morphology of MCF-7 Cells under Stress
2
Immunocytochemistry of Differentiated Cell Types
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The differentiated DE and HLCs were fixed with 3.7% formaldehyde for 30 min and permeabilized with 0.25% Triton X-100 for 10 min at 20ºC. Cells were blocked with 1% bovine serum albumin (BSA) in DPBS for 1 h prior to incubation of the cells with primary antibodies overnight at 4ºC. For induced DE cells, 1:100 diluted mouse anti-sex determining region Y-box 17 (SOX17) (ab192453, Abcam, Cambridge, UK) and 1:100 diluted rabbit anti-forehead box protein A2 (FOXA2) (ab108422, Abcam) were used. For differentiated hepatocyte-like cells, 1:200 diluted goat anti-human albumin (ALB) (sc-46293, Santa Cruz Biotechnology) and 1:200 diluted goat anti-hepatocyte nuclear factor 1-alpha (HNF-1α) (sc-6547, Santa Cruz Biotechnology) were used as primary antibodies. After washing out the unbound primary antibodies, samples were treated with CruzFluorTM 594-conjugated donkey anti-goat IgG (1:200, Santa Cruz Biotechnology), donkey anti-rabbit IgG (1:200, Santa Cruz Biotechnology), or FITC-conjugated donkey anti-mouse IgG (1:200, Santa Cruz Biotechnology) secondary antibodies for 45 min at 37ºC. The nuclei of cells were counterstained with 1 µg/ml 4',6-diamidino-2-phenylindole (DAPI) for 5 min and images were taken using an Eclipse Ti-U fluorescence microscope (Nikon Instrument).
3
3D Ovarian Cancer Cell Culture for ATR Evaluation
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The 3D cell culture system mimics the in vivo environment and serves as a unique platform to evaluate how ATR is related to in vivo ovarian cancer cell growth. Consistent with the manufacturer's protocol, the ovarian cancer cell lines Skov3 and OVCAR8 were mixed with 3D VitroGel TM (TheWell Bioscience Inc., NJ, USA) then established in 24-well plates at a density of 1× 10 4 cells/ml. Each well was covered with the same volume of cell culture medium. The experimental group received an additional treatment of VE-822 0.1 μM/ml. The plates were then placed in a 37 °C incubator with a humidified 5% CO 2 atmosphere; the covering medium was changed every 48 hours. Images of the cell spheroids were obtained with a Nikon microscope every three days. After 15 days, calcein-AM (Thermo Fisher Science) was applied to stain the tumor spheroids, and images were obtained with an Eclipse Ti-U fluorescence microscope (Nikon) equipped with a Spot RT digital camera.
4
Microfluidic Assays for Cell Imaging
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Microfluidic assays were performed by using a Bioflux 200 system (Fluxion Biosciences Inc., USA). Cells were grown in TB medium at 30°C until an OD600 of 0.5 was reached. The cells were then diluted in fresh TB until an OD600 of 0.05 was reached and flushed into the channels for 3 h at 0.5 dyn/cm2. Afterwards, cells were removed from the input well, and fresh TB medium supplemented with the respective compounds at the indicated concentrations was flushed into the channels overnight at 0.5 dyn/cm2. An exception was tyloxapol, where the medium already contained tyloxapol during the first 3 h of incubation. Imaging was performed on a Nikon Eclipse Ti-U fluorescence microscope equipped with an iXon3 897 electron-multiplying charge-coupled-device (EMCCD) camera using a 40× objective and a GFP (excitation, 470 ± 20 nm; emission, 525 ± 25 nm) filter set. Two positions per channel were imaged per strain. Quantification of whole fluorescence was performed using Fiji software.
5
Immunofluorescence Staining for Oxidative Stress and Apoptosis
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Immunofluorescence staining was performed according to standard protocols (Sun et al., 2020 (link); Li et al., 2021 (link)). The paraffin embedded heart sections and cell slides were incubated with the following primary antibodies at 4C overnight: goat anti-8-hydroxy-2′-deoxyguanosine (8-OHdG) (Sigma, Cat. # AB5830, 1:500 dilution) and rabbit anti-caspase-3 antibody (Abcam, Cat. # ab32351, 1:500 dilution). After washing, sections were incubated with donkey anti-goat IgG 647 (Abcam, Cat. # ab150135, 1:1,000 dilution) or goat anti-rabbit IgG 555 (Abcam, Cat. # ab150078, 1:1,000 dilution) at room temperature for 2 h, and counterstained with 1 μg/ml DAPI. All fluorescence images were obtained using the Nikon Eclipse Ti-U fluorescence microscope and analyzed from five randomly selected fields by ImageJ.
6
Immunofluorescence Analysis of Autophagy and Myofibroblast Markers
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After the stained cells were cultured in a 32-well plate and the intervention factors were added, the cells were fixed with 4% paraformaldehyde for 20 min and incubated with 0.5% Triton X-100 in PBST for 20 min. After blocking with 10% homologous serum of the secondary antibody for 30 min, the cells were treated with primary antibodies against LC3 (Abcam, ab192890,1:400) and α-SMA (Abcam, ab7817,1:1,400) overnight at 4°C. The following day, the cells were incubated with DyLight 488 AffiniPure goat IgG(H + L) at room temperature in the dark for 2 h. The nuclei were counterstained with 0.5 µg/mL 4′,6-diamidino-2-phenylindole for 5 min. After each step, the cells were gently washed three times with PBS for 3 min each wash. Images were obtained using an Eclipse Ti-U fluorescence microscope (×400; Nikon, Tokyo, Japan).
7
Cardiomyocyte Size Quantification using WGA
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Slides were stained with Alexa Fluor 488–conjugated wheat germ agglutinin WGA (Sigma). In brief, slides were dewaxed, rehydrated, and blocked with 3% BSA for 20 min. The slides were then incubated in WGA solubilized in PBS for 30 min at room temperature in the dark. After washing with PBS, the sections were stained with DAPI (Invitrogen) for 5 min and images were acquired using a Nikon Eclipse Ti‐U fluorescence microscope (Minato‐ku, Tokyo, Japan). The cardiomyocyte size was determined by dividing the total area by the number of cardiomyocytes using Image J software (NIH, Bethesda, MD, Unites States).
8
Aβ Localization in NPCs on Laminin
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NPCs (28,000 cells/well) were cultured and treated on laminin coated 8-chambered glass slides (ibidi USA, Madison, WI). Then, the cells were washed with PBS and fixed with absolute ethanol for 30 min at 4 °C. After washing with PBS, nuclei were stained with DRAQ5 for 5 min followed by PBS wash. Slides were mounted using ProLong Gold antifade reagent (Life Technologies). Green fluorescence (originating from Aβ HiLyte Alexa Fluor488) and red fluorescence (from DRAQ5) were acquired directly using a Nikon Eclipse Ti-U fluorescence microscope.
9
Immunofluorescence Staining of Cardiomyocytes
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Experimental primary cardiomyocytes in 48-orifice-plates were fixed using 4% paraformaldehyde, permeabilized using 0.5%Triton X-100, blocked using 4% goat serum, and then incubated with primary antibodies against SM α-actin (α-SMA, ab5694), and connexin 43 (ab11370) at 4°C overnight. After incubation with DyLight 488 and 594 AffiniPure Goat IgG (H + L) for 1 h at 37°C, the cardiomyocytes were counterstained with 0.1 μg/mL 4′,6-diamidino-2-phenylindole (DAPI) (P36941; Invitrogen) for 3 min and images were obtained using a Nikon Eclipse Ti-U fluorescence microscope (×400 or ×200).
10
3D Cell Culture Assay for Myc Effects on Osteosarcoma
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In order to simulate the in vivo environment, a three-dimensional (3D) cell culture assay was used to evaluate the effect of Myc on osteosarcoma cell growth. According to the manufacturer’s protocol, we mixed the hydrogel with the osteosarcoma cells at a density of 1 × 104 cells/ml, then seeded them in a 24-well VitroGel 3D cell culture plate (The Well Bioscience, Newark, NJ, USA) covered with different cell culture media (with or without 10 μM 10058-F4). The plate was placed in an incubator and the covering medium was changed every 48 h. Every 3 days, spheroids were selected based on their size, volume, and morphology, and imaged by microscope equipped with a digital camera. A cell culture medium containing 0.25 μM calcein AM (Thermo Fisher Science) was applied 15 days later to cover the hydrogel. Spheroids were imaged 15 min after incubation, with an Eclipse Ti-U fluorescence microscope (Nikon) equipped with a SPOT real-time (RT) digital camera. The diameter of spheroids was measured three times using ImageJ software as previously described (https://imagej.nih.gov ).15 (link),20 (link)
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