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. 2021 Aug 26;188(2):683–702. doi: 10.1093/plphys/kiab349

Figure 6.

Figure 6

Super-resolution imaging applications in plants. A–E, Multicolor SIM fluorescent localization of (A) Alexa Fluor 647-labeled MTs, (B) DAPI-labeled chromosomes, (C) Alexa Fluor 488-labeled γ-tubulin, and (D) Alexa Fluor 546-labeled pERK species in a barley anaphase root tip cell. E, Magnified overlay of A and B. F and G, Immunofluorescence SIM colocalization of cortical MTs (magenta), PLDα1-YFP (green), and clathrin (blue) in Arabidopsis root cells. Boxed area in (F) is shown magnified in (G). Arrows indicate PLDα1-YFP spot-like structures in close contact with cortical MTs and individual clathrin-coated pits and vesicles. H, Live SIM imaging of GFP-MBD labeled cortical MTs of alfalfa pavement leaf cells after application of perfluorodecalin. I and J, PALM visualization of the MT-associated protein MAP65-2 fused to (I) mEos3.2 and (J) the respective HiLO widefield image. Bar = 10 µm (H), 5 µm (A–E), and 1 µm (F, G, I, and J). Imaging: Oil Plan-Apochromat 63×/1.4 NA (A–I), 405 nm at 6.5% for DAPI excitation (A–E) and 5.0% for photoconversion of mEos3.2 (I–J), 488 nm at 3.5% (A–E), 2% (F–H) and 2% for excitation of non-converted mEos3.2 (I–J), 561 nm at 2.8% (A–E), 2% (F and G) and 100% for excitation of photoconverted mEos3.2 (I–J), 642 nm at 3% (A–E), and 2% (F and G), signal detection with BP420–480+LP750 for DAPI (A–E), BP 495–550 + LP 750 for Alexa Fluor 488 (A–G) and for GFP (H), BP 570–620 + LP 750 for Alexa Fluor 546 (A–G), LP 655 for Alexa Fluor 647 (A–G), a combination of BP 420–480 + BP 570–640 + LP 740 for PALM visualization (I–J), five rotations, and five phase shifts for SIM (A–H).