Isolated cardiomyocytes were placed in a polycarbonate recording chamber (Warner Instruments, Hamden, CT) on a confocal microscope stage, and cells were allowed to settle and spontaneously attach to the bottom of the recording chamber for 10 min. Cells were imaged using an inverted laser-scanning confocal microscope (Nikon Eclipse TE2000-U, Tokyo, Japan) with a 60x/1.4 oil-immersion objective (Nikon). Settings of the confocal microscope were consistent in all experimental groups. Fluorescent images were acquired using the EZ-C1 2.10 software (Nikon) and data were analyzed off-line with the MetaMorph 6.2 software (Universal Imaging, West Chester, PA). Results were expressed as percent change in fluorescence intensity relative to baseline (F0; where baseline = 100%).
Ez c1 2
Manufactured by Nikon
The EZ-C1 2.10 software is a imaging analysis tool developed by Nikon for their line of confocal microscopes. The software provides users with the ability to capture, process, and analyze images obtained from Nikon's confocal systems.
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Lab products found in correlation
2 protocols using ez c1 2
1
Confocal Microscopy of Cardiomyocytes
2
Quantifying Nitric Oxide in HTR8/SVneo Cells
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To determine the nitric oxide levels, HTR8/SVneo was cultured on the various matrices. HTR8/SVneo was incubated for 30 min with 4-amino-5-methylamino-2′7′diaminofluorescein (DAF; 10 μM; Invitrogen, Carlsbad, CA) for the determination of nitric oxide (NO) levels. The fluorescent stain was removed, and the cells were allowed to stabilize for 30 min. The slides were then sealed with an aqueous mounting media and analyzed immediately by confocal microscopy (Eclipse TE2000-U; Nikon). Images were recorded with EZ-C1 2.10 software (Nikon). Images were captured using 488 nm excitation for and >519 nm emission, respectively. Fluorescence intensity was measured by determining pixel intensity of each cell, and only viable cells were counted.
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