Extended Data Fig. 2: Cell-wide, spatiotemporally high-resolution, sustainable imaging of ER network rearrangement.
From: A highly photostable and bright green fluorescent protein
The three neighboring HeLa cells expressing er-(n2)oxStayGold(c4) (Fig. 4a–d) were analyzed. Scale bars, 5 μm. a, Magnified SIM and binarized images at t = 0 min (First frame) and t = 6 min (Last frame) of a peripheral region of the uppermost cell. Compare with Supplementary Fig. 11i. The preservation of 3D-SIM image quality in the last frame was confirmed in all six-minute experiments that employed er-(n2)oxStayGold(c4). Shown is a representative of n = 9 independent transfections, including experiments #1, #2, #3, and #4 (Supplementary Fig. 11a-d). b, Automatic quantification of ER network rearrangement. See Methods (Automatic quantification of ER network rearrangement). Magnifications of the boxed region are shown (bottom). Considering that the diameter of an ER tubule spanned several pixels, we reasoned that the sub-block size was optimal for the efficient detection of target movement.
