Fig. 4.

Overlaid multi-focus fluorescence and transmission images of living budding yeast (S. cerevisiae) cells expressing green fluorescent protein (GFP) tagged with a rigid linker to septin. Septin incorporates into the hourglass/ring structure at the bud neck during cell division. 3D protein organization analysis of the cells (i) and (ii), marked with white rectangles, are shown in Fig. 5. Multi-focus imaging allows screening of many cells simultaneously, and allows stable imaging over long time periods. The focus step is 250nm between planes and the lateral size of each focal slice is 33 × 33 μm. See Media 1 ^{(88.6KB, MOV)} for a 3D visualization of the septin rings.