All widefield images were captured on an inverted Nikon TE2000-E widefield microscope with Volocity Acquisition software (PerkinElmer). Following objective lenses were used: 4× objective lens (Plan Fluor 4×/NA 0.13), 10× objective lens (UPLSAPO 10×/NA 0.30 DIC), and 20× objective lens (UPLSAPO 20×/NA 0.45). Confocal laser scanning microscope imaging used a Fluoview Spectral FV1200 CSLM based on Olympus IX83 motorized inverted microscope CSLM. Following objective lenses were used: 60× objective lens (UPLSAPO 60×/NA 1.4) and a 30× objective (UPLSAPO 30×/NA 1.05 Silicone Immersion). Spinning disc confocal microscopy imaging was performed on a PerkinElmer Spinning Disk Confocal Microscope based on a Nikon TE 2000-U Eclipse motorized inverted microscope with DIC optics. The following objective lens was used: 60× (Plan Apo 60×/NA 1.40). Volocity software (PerkinElmer) was used for Acquisition.
Te 2000 u eclipse
Manufactured by Nikon
The Nikon TE 2000-U Eclipse is a research-grade inverted microscope designed for advanced cell and tissue culture applications. It features a stable and vibration-resistant frame, providing a reliable platform for high-resolution imaging. The TE 2000-U Eclipse supports a range of observation techniques, including phase contrast, differential interference contrast (DIC), and fluorescence microscopy, allowing researchers to explore a variety of biological samples.
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2 protocols using te 2000 u eclipse
1
Widefield, Confocal, and Spinning Disk Microscopy
2
Immunofluorescent Staining of Tight Junction Proteins
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Confluent cells were fixed in 4% paraformaldehyde in PBS, pH 7.2, for 10 min at 37°C, then permeabilized in 0.1% Triton X-100 (Sigma-Aldrich; Merck KGaA) in PBS for 5 min at room temperature; rinsed three times in PBS, and blocked for 30 min in PBS with 10% goat serum (Sigma-Aldrich; Merck KGaA) at 37°C. Primary antibody staining was performed at 4°C overnight with the following antibodies at a 1:50 dilution: Anti-ZO-1 and anti-ZONAB. The sections were then incubated with an Alexa Fluor® 594-conjugated goat anti-mouse secondary antibody at a 1:100 dilution (cat. no. SA00006-3; ProteinTech Group, Inc.) for 2 h at room temperature in the dark. Following DAPI staining at 37°C for 10 min, cells were imaged using an inverted fluorescence microscope (TE2000U Eclipse; Nikon Corporation).
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