Super-resolution PALM imaging reveals clustering of Swi4-, Mbp1-, and Swi6-mEos3.2 in fixed budding yeast cells.
(A–C) PALM imaging of from Swi4- (A), Mbp1- (B), and Swi6-mEos3.2 (C). For all panels: top panels in A–C: Composite images of the phase contrast and PALM detection images with, left to right, the full FOV and zoomed images of a small, medium, and large cell (indicated by the yellow circles in the full FOV image). Scale bar for the full FOV is 5 µm for Swi4-mEos3.2 and Swi6-mEos3.2 and 10 µm for Mbp1-mEos3.2. Scale bars for the zoomed composite images are 1 µm. Detection image outputs (pink dots; one for each detection) were obtained with the Thunderstorm plugin from ImageJ and were not filtered for blinking. High-intensity purple in the image corresponds to out-of-focus beads. Lower panels in A–C: Zoomed molecular images of the small, medium, and large cells in the composite images in the top panels of A–C. Molecular images were created as described in the text and corrected for blinking. Scale bars in panels, left to right, are 0.15 µm, 0.24 µm, and 0.3 µm for Swi4-mEos3.2, 0.21 µm, 0.21 µm, and 0.21 µm for Mbp1-mEos3.2 and 0.15 µm, 0.15 µm, and 0.27 µm for Swi6-mEos3.2.