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Simultaneous Zn2+ tracking in multiple organelles using super-resolution morphology-correlated organelle identification in living cells
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Simultaneous Zn2+ tracking in multiple organelles using super-resolution morphology-correlated organelle identification in living cells
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Journal Article
Simultaneous Zn2+ tracking in multiple organelles using super-resolution morphology-correlated organelle identification in living cells
2021
Overview
Zn
2+
plays important roles in metabolism and signaling regulation. Subcellular Zn
2+
compartmentalization is essential for organelle functions and cell biology, but there is currently no method to determine Zn
2+
signaling relationships among more than two different organelles with one probe. Here, we report simultaneous Zn
2+
tracking in multiple organelles (Zn-STIMO), a method that uses structured illumination microscopy (SIM) and a single Zn
2+
fluorescent probe, allowing super-resolution morphology-correlated organelle identification in living cells. To guarantee SIM imaging quality for organelle identification, we develop a new turn-on Zn
2+
fluorescent probe, NapBu-BPEA, by regulating the lipophilicity of naphthalimide-derived Zn
2+
probes to make it accumulate in multiple organelles except the nucleus. Zn-STIMO with this probe shows that CCCP-induced mitophagy in HeLa cells is associated with labile Zn
2+
enhancement. Therefore, direct organelle identification supported by SIM imaging makes Zn-STIMO a reliable method to determine labile Zn
2+
dynamics in various organelles with one probe. Finally, SIM imaging of pluripotent stem cell-derived organoids with NapBu-BPEA demonstrates the potential of super-resolution morphology-correlated organelle identification to track biospecies and events in specific organelles within organoids.
Subcellular Zn
2+
compartmentalisation is essential for cell biology. Here the authors make a turn-on fluorescent Zn
2+
probe that localises to multiple organelles, and correlate its location using organelle morphology derived from structured illumination microscopy.
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