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. Author manuscript; available in PMC: 2023 Feb 7.
Published in final edited form as: Curr Opin Chem Biol. 2021 Oct 27;65:154–162. doi: 10.1016/j.cbpa.2021.09.003

Figure 2. Lipid-focused strategies to image organelles and/or study their trafficking.

Figure 2

(a) (i) Cer-TCO, which localizes in a temperature-dependent way to the Golgi or ER and reacts and (ii) the environment-dependent spirocyclization equilibria of SiR and HMSiR that effectively extends imaging times by ‘hiding’ a reservoir of dark-state dyes in the membrane. (b) in cellula with tetrazine-containing fluorophores such as SiR and HMSiR to enable STED and SMLM, respectively. (c) Biosynthetic incorporation of azido-choline into phospholipids followed by coincubation with a click-compatible dye fused to an organelle-targeting small molecule results in selective localization of the labeled lipid to the organelle of interest. (d) Incorporation of oxo-TCO handle viaPLD-mediated transphosphatidylation, followed by fluorogenic Tz-BODIPY results in real-time visualization of PLD activity. BODIPY, boron dipyrromethene difluoride; ER, endoplasmic reticulum; PLD, phospholipase D; SMLM, single-molecule localization microscopy; STED, stimulated emission depletion; HMSiR, hydroxymethyl silicon rhodamine; SiR, silicon rhodamine; DBCO, dibenzocyclooctyne; TCO, trans-cyclooctyne.