Nitric Oxide production in the cell culture media was determined by measuring nitrite levels with a Siever’s NO analyzer. Nitric oxide levels in HPMEC were also determined using DAF-DA as per manufacture instructions (Life Sciences). Briefly, cells were loaded with DAF-DA (5 μM) for 20 min and washed 3 times with PBS and incubated for another 30 min in phenol-free media. Fluorescence was determined using a Nikon TE-200 epifluorescence microscope. Images were captured with a Rolera EMC2 camera (Q-Imaging, BC, Canada) with Velocity software (PerkinElmer, Lexington, MA).
Te200 epifluorescence microscope
Manufactured by Nikon
The Nikon TE200 is an epifluorescence microscope designed for laboratory use. It provides the core function of illuminating and observing fluorescently-labeled samples.
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Lab products found in correlation
3 protocols using te200 epifluorescence microscope
1
Quantifying Nitric Oxide in Cell Cultures
2
Cell Viability Assay by Propidium Iodide
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Cells seeded onto 35-mm culture dishes received vehicle or drug treatment directly to their culture medium and were then incubated for 48 h (5% CO2, 37 °C). The media was replaced, and propidium iodide (10 μM) added to test for viability as a function of dye exclusion. After a 10-min incubation at room temperature, cell viability within each dish was assessed based on the number of propidium iodide–positive cells manually counted across three fields of view under the x10 microscope objective attached to a Nikon TE200 epifluorescence microscope (fluorescence observed through a 530 nm excitation/620 nm emission filter). Counts were made by an investigator blinded to the treatment regimen.
3
Epifluorescence microscopy of cells
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