Super-resolution PALM imaging of Whi5-mEos3.2 in SC + 2% glucose, Swi4-mEos3.2, Mbp1-mEos3.2, and Swi6-mEos3.2 in SC + 2% glycerol and NLS-mEos3.2-NLS in SC + 2% glucose. (A) PALM imaging of Whi5-mEos3.2. Top panels: Composite phase contrast image and the detection image (red dots; one for each detection) output produced with Thunderstorm. The detection image is not filtered for blinking. Saturated red intensity in the image corresponds to out-of-focus beads or compromised cells. Left to right are the full FOV and zoomed cells 1 (small), 2 (medium), and 3 (large) described below are indicated by yellow circles and numbers. Cells were grown in SC + 2% glucose. Scale bar is 5 µm for full FOV and 1 µm for each of the zoomed images. Bottom panels: Molecular images of the nuclei of indicated cells 1–3, respectively, left to right (from yellow squares in top panels). Molecular images were created as described in the text and corrected for blinking. Scale bars, 0.3 µm. (B) Molecular images of cells in SC + 2% glycerol medium expressing, left to right, Swi4-mEos3.2, Mbp1-mEos3.2, and Swi6-mEos3.2, respectively. Scale bars are 0.18, 0.24, and 0.3 µm, left to right, respectively. (C) PALM imaging of the NLS-mEos3.2-NLS control. Top panels: Composite phase contrast images and the detection images (red dots; one for each detection) output was produced with Thunderstorm. The detection image is not filtered for blinking. Images from left to right correspond to the full FOV, and cells 1–3, respectively. Scale bar in full FOV is 5 µm, and 1 µm for the zoomed cells 1–3. Bottom panels: Molecular images of the nuclei of indicated cells 1–3, respectively, left to right (from yellow squares in top panels). Molecular images were created as described in the text and corrected for blinking. Scale bars, 0.3 µm.