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Eclipse t1

Manufactured by Nikon
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Sourced in Japan

The Nikon Eclipse T1 is a microscope designed for laboratory use. It features a compact and ergonomic design, providing a stable and consistent platform for various imaging and analytical applications.

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Lab products found in correlation

Exoglow protein ev labeling kit, by System Biosciences (2 mentions) Cfi apo tirf 100x na 1, by Nikon (2 mentions) Dmi6000b inverted microscope, by Leica (1 mentions) Femtojet, by Leica (1 mentions) Alexa fluor 647 conjugated dextran, by Thermo Fisher Scientific (1 mentions) Bovine pituitary extract, by Absin (1 mentions) Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions) Alexa fluor 647 conjugated transferrin, by Thermo Fisher Scientific (1 mentions) X tremegene hp dna transfection reagent, by Roche (1 mentions) 8 well chamber slide, by Ibidi (1 mentions)

Close spelling variants of this product

Eclipse T1 inverted optical microscope Nikon Eclipse T1 microscope Nikon Eclipse T1 confocal microscope

6 protocols using eclipse t1

1

Visualizing ER dynamics in U2OS cells

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U2OS cells were seeded on a glass bottom dish, well size 14 mm (Cellvis) and transfected with 0.1 μg of a vector encoding for EGFP-KDEL 48 hr before the experiment. Subsequently, a mixture of Alexa Fluor 647 conjugated dextran (0.5 mg/ml, MW 11 kDa) (Thermo Fischer Scientific) and GTP or GTPγS (10 mM) in permeabilization buffer was injected into the cytoplasm of cells using an Eppendorf FemtoJet and InjectMan Ni2 microinjection unit at a Leica DMI6000B inverted microscope equipped with a 20X, 0.4 NA, Ph1 HCX PlanFluotuar LD (long distance) objective. ER dynamics in microinjected cells was imaged for 1 min at a frame rate of 1/s using a Visitron spinning disk Nikon Eclipse T1 microscope with a 100X, 1.49 CFI, Apo TIRF objective. Image analysis was performed with ImageJ. Brightness and contrast were adjusted across the images. From each cell, 1–2 ROIs of 100 μm2 in the peripheral ER were selected for manual analysis of successful or unsuccessful membrane attachments.
Pawar S., Ungricht R., Tiefenboeck P., Leroux J.C, & Kutay U. (2017). Efficient protein targeting to the inner nuclear membrane requires Atlastin-dependent maintenance of ER topology. eLife, 6, e28202.
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2

Visualizing EV Uptake by Neurons

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An ExoGlow-protein EV labeling kit (System Biosciences, Palo Alto, CA) was used for live cell EV imaging. Collected EVs were incubated for 20 minutes with the labeling dye (1:500) at 37 °C to induce vesicular protein conjugation with the dye molecules. Then the solution was treated with the ExoQuick-TC solution and incubated for 2 hours at 4 °C, followed by centrifugation at 10,000 rcf for 10 minutes to remove unlabeled reagent molecules and collect labeled EVs only. The labeled EVs were resuspended in neuronal maintenance medium and added to neurons 72 hours post-differentiation at various concentrations. A live cell imaging microscope (Eclipse T1, Nikon) was used to image fluorescently labeled EV uptake by the cultured neurons. In these experiments, images were collected at multiple locations from examined culture wells every 5 minutes for 2 hours. Additionally, neurons treated with unlabeled EVs were fixed after 2 hours of treatment and immunostained with a mouse anti-CD81 antibody (1:100, Thermo-Fisher) to quantify the relative amount of internalized EVs based on fluorescence intensity. CD81 labeling was coupled with fluorescently-labelled phalloidin (for F-actin visualization; 1:200, Invitrogen) to confirm the correct positioning of CD81 expression relative to the cell cytoskeleton.
Chun C., Smith A.S., Kim H., Kamenz D.S., Lee J.H., Lee J.B., Mack D.L., Bothwell M., Clelland C.D, & Kim D.H. (2021). Astrocyte-derived Extracellular Vesicles Enhance the Survival and Electrophysiological Function of Human Cortical Neurons in vitro. Biomaterials, 271, 120700.
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3

Culturing Normal Prostatic Epithelial Cells

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Normal prostatic epithelial RWPE-1 and HPr-1 cells (Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai, China) were cultured in complete keratinocyte serum-free medium (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany), supplemented with 1% penicillin/streptomycin/amphotericin B, 50 μg/ml bovine pituitary extract (Absin Bioscience, Inc., Shanghai, China) and 5 ng/ml epidermal growth factor (Thermo Fisher Scientific, Inc., Waltham, MA, USA). The cells were incubated at 37°C in an atmosphere containing 5% CO2. When cells reached 70-80% confluence they were washed with PBS and detached using 0.25% trypsin/0.2% EDTA. Cells were viewed under a light microscope (magnification, ×200; Eclipse T1, Nikon Corporation, Tokyo, Japan), and subsequently suspended at a concentration of 1×106 cells/ml.
Ye C., Cai Y., Cai Q., Yuan S., Huang F., Yang X., He S., Li Z., Wang Y., Yang D, & Li Z. (2018). High glucose induces the proliferation of prostatic cells via downregulating MRE11. International Journal of Molecular Medicine, 41(6), 3105-3114.
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4

Transferrin Uptake Quantification

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Cells were incubated with Alexa Fluor 647-conjugated transferrin (T23366, Life Technologies, distributed by ThermoFisher Scientific, Waltham, USA) for 15 min, washed with PBS and fixed with paraformaldehyde (PFA). Images were acquired on a confocal microscope (Nikon Eclipse T1, Nikon Tokyo, Japan) with spinning disk (3I, Denver, CO, USA) with a 100x oil objective (Nikon CFI Apo TIRF 100x NA 1.4). z-stack images in a total range of 10 µm were acquired and maximum projections obtained with ImageJ. The quantification of the fluorescence signal was performed in ImageJ by measuring the mean grey value for the transferrin channel (far red) per pixel in the area of the cells.
Hohendahl A., Talledge N., Galli V., Shen P.S., Humbert F., De Camilli P., Frost A, & Roux A. (2017). Structural inhibition of dynamin-mediated membrane fission by endophilin. eLife, 6, e26856.
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5

Dynamin2 and Endophilin A2 Interactions

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Genome-edited SK-MEL-2 cells expressing the endogenous dynamin2 with a GFP tag (kind gift from David Drubin lab, UC Berkeley, USA; [Doyon et al., 2011 (link)]) were transfected using 0.5 µg of plasmids containing human endophilin A2 with C-terminal TagRFP-T under the control of the CMV promoter with X-tremeGENE HP DNA Transfection Reagent (06366236001, Roche Diagnostics GmbH, Mannheim, Germany) according to the manufacturer’s protocol. The endophilin plasmid was kindly provided by Emmanuel Boucrot, University College London, UK. For the recovery experiments, human dynamin2 cherry (kindly provided by Christien Merrifield, LEBS, Gif-Sur-Yvette, France) was overexpressed together with a GFP version of human endophilinA2 (kindly provided by Emmanuel Boucrot [Boucrot et al., 2015 (link)]). Images of the cells were acquired on a microscope (Nikon Eclipse T1, Nikon Tokyo, Japan) with a 100x oil objective (Nikon CFI Apo TIRF 100x NA 1.4) using total internal reflection fluorescence (TIRF).
Hohendahl A., Talledge N., Galli V., Shen P.S., Humbert F., De Camilli P., Frost A, & Roux A. (2017). Structural inhibition of dynamin-mediated membrane fission by endophilin. eLife, 6, e26856.
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6

Live/Dead Viability Assessment of Bioprinted Scaffolds

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LIVE/DEAD® Viability/Cytotoxicity assay was tested on scaffolds (n ≥ 3) after 1 day of bioprinting and after 15 days of compression loading to assess the impact of bioprinting and compression loading on cell viability after bioprinting and after 2 weeks of compression loading. Briefly, scaffolds were incubated with 2 μM Calcein AM and 4 μM ethidium homodimer for 40 min 37°C and 5% CO2. Then, scaffolds were washed twice with pre-warmed PBS and transferred to 8-well chamber slides (Ibidi GmbH, Germany) for imaging using a confocal microscope (Visitron Spinning Disc, Nikon Eclipse T1). For each scaffold, 6 distinct regions were imaged. Cell viability was calculated using ImageJ (National Institutes of Health, United States of America) as the ratio of the number of living cells to the total number of cells. Cell density (cells/mm2) was estimated using ImageJ as number of living cells per scaffold area.
de Leeuw A.M., Graf R., Lim P.J., Zhang J., Schädli G.N., Peterhans S., Rohrbach M., Giunta C., Rüger M., Rubert M, & Müller R. (2024). Physiological cell bioprinting density in human bone-derived cell-laden scaffolds enhances matrix mineralization rate and stiffness under dynamic loading. Frontiers in Bioengineering and Biotechnology, 12, 1310289.
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