Single pole photobleach and recovery.

<p>A) 1. Initial fluorescence, 2. Image immediately after shutter (red dashed area) removed, 3. Image after 15 min recovery, 4. Image after 25 min recovery. B) Intensity profiles along yellow dashed line in A1 showing fast recovery of the brighter, sheltered pole.</p

Whole cell recovery after photobleaching.

<p>A) Fluorescence before photobleaching (pre-bleach). B) Fluorescence after 30 min recovery (post recovery). C) Combined image (Green: pre-bleach, Red: post recovery). Overlaid cell boundaries have been added to guide the eye. D) Initial fluorescence of cells 1, 2, and 3 in Panels A. E) Fluorescence of cells 1, 2, and 3 after photobleaching and 30 min. recovery showing fast recovery of one pole and slow recovery of other pole. F) and G) Fluorescence profiles showing asymmetric recovery after 30 min and 1 hour, respectively.</p

Localization imaging of the nascent new pole.

<p>A) Representative integrated image and filmstrip showing escape of molecules from the brighter old pole and diffusion toward the new pole region. B-C) Localization images of the propensity of Tsr-Venus to reside in the entire cell. The sharp bright dots show an artefactually small localization of the bright stable old pole cluster. At the opposite poles, the areas where the new pole forms show no indication of cluster formational though bulk fluorescence imaging (insets).</p