Cells were imaged in an enclosed 37°C, 5% CO2 incubation chamber. Images were taken on the 561 nm (paxillin TMR) and the 642 nm (beads) channels with a 500 ms exposure time every 5 seconds for 200 frames using a Nikon Ti Total Internal Reflection Fluorescence (TIRF) microscope using a 100× objective with 1.5× additional magnification factor with a Hamamatsu ORCA-D2 CCD camera (Hamamatsu Corporation, Bridgewater. NJ, USA, final resolution: 43 nm/pixel). To obtain a bead image from the undeformed substrate, cells were removed from the substrate by injecting a high dose (0.5%) of 5 mL Trypsin/EDTA (Invitrogen) for 30 min.
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Imaging Substrate Deformation in Cells
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Other organizations : The University of Texas Southwestern Medical Center, Harvard University, University of California, San Diego
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Variable analysis
independent variables
- Removal of cells from the substrate by injecting a high dose (0.5%) of 5 mL Trypsin/EDTA (Invitrogen) for 30 min
- Bead image from the undeformed substrate
- Cells were imaged in an enclosed 37°C, 5% CO2 incubation chamber
- Images were taken on the 561 nm (paxillin TMR) and the 642 nm (beads) channels with a 500 ms exposure time every 5 seconds for 200 frames
- Nikon Ti Total Internal Reflection Fluorescence (TIRF) microscope using a 100× objective with 1.5× additional magnification factor with a Hamamatsu ORCA-D2 CCD camera (Hamamatsu Corporation, Bridgewater. NJ, USA, final resolution: 43 nm/pixel)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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