PALM imaging process using ultrafast data acquisition of mEos3.2-paxillin and reconstructed images, which are shown in Fig. 4, A and C. A data acquisition rate of 1 kHz for a period of 10 s (10,000 frames) was employed for 640 × 640 pixels (35.3 × 35.3 µm2), covering most of the basal PM area, including many FAs. Fluorescent spot clusters found in the initial 30 frames (0.03 s) of the data acquisition sequence are due to the presence of mEos3.2 pre-photoconverted during the search for the cells suitable for PALM imaging, and thus they were excluded from the image reconstruction process.
