Prepared 96-well plates were imaged on a Nikon Ti-E inverted microscope with a stage-top incubator (37°C, 5% CO2). Coordinates within each well of the 96-well plate were imaged at 6 minute increments which were automated by the Nikon Elements AR software. Images were captured using an Andor Zyla 5.5 scMOS camera and a 20x/0.75 NA objective. Chroma #49001 (ET-CFP) and #49003 (ET-YFP) excitation/emission filter cubes were used for mTurquoise2 and YPet measurements, respectively. Further details are described in (Pargett et al., 2017 (link)). Coordinates of each acquisition area were saved for future imaging of immunostaining experiments.
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Time-lapse Imaging of Cell Signaling
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Other organizations : University of California, Davis
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Variable analysis
independent variables
- None explicitly mentioned
- Measurements of mTurquoise2 and YPet fluorescence
- Temperature (37°C)
- CO2 concentration (5%)
- Coordinates within each well of the 96-well plate
- Acquisition time interval (6 minutes)
- None explicitly mentioned
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