Eclipse te2000 e confocal

Manufactured by Nikon

The Nikon Eclipse TE2000-E is a confocal microscope designed for advanced imaging applications. It features a modular design that allows for customization to meet specific research needs. The core function of the Eclipse TE2000-E is to provide high-resolution, three-dimensional imaging capabilities using laser-scanning technology.

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Lab products found in correlation

Axio observer z1, by Zeiss (3 mentions) Alexa fluor 488 goat anti rabbit igg secondary antibody, by Thermo Fisher Scientific (1 mentions) Lipofectamine ltx, by Thermo Fisher Scientific (1 mentions) Accuri c6 cytometer, by BD (1 mentions)

Close spelling variants of this product

Nikon Eclipse TE2000-E Confocal microscope Eclipse TE2000-E inverse confocal laser scanning microscope Sweptfield/Eclipse/TE2000-E confocal microscope Eclipse TE2000-E confocal laser scanning microscope Eclipse TE2000-E Eclipse TE2000 TE2000-E TE2000

4 protocols using eclipse te2000 e confocal

Cell Culture, Transfection, and Imaging

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HEK293T, HuH-7 and Caco-2 cells were maintained in DMEM supplemented with 10% FBS at 37°C in 5% CO2. Transient transfections were performed using Lipofectamine LTX (Invitrogen - Catalog #15338100) and GeneJuice (Novagen -Catalog #70967-3) according to manufacturer's instructions. Lentiviral infections of Caco-2 cells were performed as detailed in Langlois et al.⁷² (link) All transfections with multiple plasmids were performed using an equal amount of overall plasmid DNA. Differences in the amount of DNA from experimental constructs were filled with an empty construct.
FACS analysis was performed on a Accuri C6 cytometer (BD) after transfection/infection of the cells, at the times indicated in the figure legends.
Cell visualization was performed on an Eclipse TE2000-E confocal (Nikon) confocal microscope (Fig. 5), or on a Axio Observer Z1 (Zeiss) inverted microscope (Fig. S4). Transiently transfected cells were replated on coverslips after 24 h from transfection, and then fixed by 4% Formaldehyde and stained at 48 hr. Immunocgemistry for EGFP was carried out using a rabbit anti-GFP antibody (1:500; Molecular Probes – Catalog #A11122) and a AlexaFluor488 goat anti-rabbit IgG secondary antibody (1:500; Molecular Probes – Catalog #A11008).

Severi F, & Conticello S.G. (2015). Flow-cytometric visualization of C>U mRNA editing reveals the dynamics of the process in live cells. RNA Biology, 12(4), 389-397.

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Immunofluorescence Microscopy of Transfected Cells

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Cells transfected with the pMT-Hth2AGFPUbx bicistronic expression vector were induced for 4 h as described above, and fixed using 4 % formaldehyde (Sigma-Aldrich F8775) in PBS for 10 min at 23 °C in an Eppendorf Thermomixer at 700 rpm. Transfected (GFP+) and non-transfected (GFP−) cells were sorted as described above. Sorted cells were seeded onto 1 × 0.8 cm coverslips in the wells of a 12-well plate each containing 1 ml PBS. Cells were allowed to adhere for 30 min. The PBS was then replaced by 1 ml PTX (PBS/0.5 % Triton X-100) and cells were incubated at room temperature for 30 min. Coverslips were then transferred to microfuge tubes containing mouse anti-Exd B11M monoclonal antibody (1 ml, 1:5 in PTX; [56 (link)]) and incubated at 4 °C overnight. Coverslips were then washed three times in PTX then incubated with goat anti-mouse Alexa 568 secondary antibody (1 ml, 1:500, Invitrogen) and 1 µg ml⁻¹ DAPI in PTX at room temperature for 2 h. Coverslips were then washed three times in PTX and mounted in Citifluor AF1 Glycerol/PBS solution (Agar Scientific) for immunofluorescence microscopy (Nikon Eclipse TE2000-E confocal).

Beh C.Y., El-Sharnouby S., Chatzipli A., Russell S., Choo S.W, & White R. (2016). Roles of cofactors and chromatin accessibility in Hox protein target specificity. Epigenetics & Chromatin, 9, 1.

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Confocal Microscopy Imaging Protocol

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Samples were imaged on a laser point-scanning confocal microscope (Nikon Eclipse TE2000-E Confocal). Z stacks were acquired with a 20×/0.6 oil lens, using a 488 nm laser paired with a 515/30 bandpass, a 546 nm laser with a 590/50 bandpass, and 647 nm laser with a 650 long pass filter. Fluorophores were excited and imaged sequentially with each laser and filter combination to minimize crosstalk with 1,024 pixel resolution and saved as 8-bit images.

Corliss B.A., Delalio L.J., Stevenson Keller TC I.V., Keller A.S., Keller D.A., Corliss B.H., Beers J.M., Peirce S.M, & Isakson B.E. (2019). Vascular Expression of Hemoglobin Alpha in Antarctic Icefish Supports Iron Limitation as Novel Evolutionary Driver. Frontiers in Physiology, 10, 1389.

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Quantifying Bacterial Adhesion on PLA and Sapphire

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In order to visually compare adhesion across samples after biopanning against PLA, 0.5 x 0.5 cm PLA coupons and sapphire pieces were incubated with induced 5 mL cultures of the naive library, sorting rounds, or negative control (which consists of a library member with an empty eCPX scaffold, i.e. no peptide). Cultures were prepared from freezer stocks grown overnight in LB+Cm. Fresh LB was inoculated 1:50 with the overnight stocks and brought to an OD of 0.5-0.6, then induced overnight with 0.4% arabinose as described above. PLA and sapphire pieces were incubated for 10 min with each culture at 230 rpm, then washed for an additional 10 min in fresh culture tubes containing 4 ml of PBS 1% Tween at 330 rpm to remove weakly bound cells. A final dip in 4 ml of fresh PBS ensured any unbound bacteria in the wash solution would be completely removed. Material samples were placed on microscope slides and visualized using an epifluorescent microscope (Nikon Eclipse TE2000-E confocal) at 40X and a red fluorescent filter (AT-TRITC/CY3). Samples were prepared independently in triplicate and ten images were captured from each sample and analyzed for the number of red fluorescent pixels using ImageJ [27 (link)]. Student’s T-tests were used to statistically verify differences in fluorescent pixel density.

Stellwagen S.D., Sarkes D.A., Adams B.L., Hunt M.A., Renberg R.L., Hurley M.M, & Stratis-Cullum D.N. (2019). The next generation of biopanning: next gen sequencing improves analysis of bacterial display libraries. BMC Biotechnology, 19, 100.

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