Abstract
Single-molecule localization microscopy (SMLM) is a rapidly evolving technique to resolve subcellular structures and single-molecule dynamics at the nanoscale. Here, we employ conventional BODIPY conjugates for live-cell SMLM via their previously reported red-shifted ground-state dimers (DII), which transiently form through bi-molecular encounters and emit bright single-molecule fluorescence. We employ the versatility of DII-state SMLM to resolve the nanoscopic spatial regulation and dynamics of single fatty acid analogs (FAas) and lipid droplets (LDs) in living yeast and mammalian cells with two colors. In fed cells, FAas localize to the endoplasmic reticulum and LDs of ~125 nm diameter. Upon fasting, however, FAas form dense, non-LD clusters of ~100 nm diameter at the plasma membrane and transition from free diffusion to confined immobilization. Our reported SMLM capability of conventional BODIPY conjugates is further demonstrated by imaging lysosomes in mammalian cells and enables simple and versatile live-cell imaging of sub-cellular structures at the nanoscale.
| Original language | English (US) |
|---|---|
| Article number | 3400 |
| Journal | Nature Communications |
| Volume | 10 |
| Issue number | 1 |
| DOIs | |
| State | Published - Dec 1 2019 |
Bibliographical note
Funding Information:We thank Doug Mashek and his lab for providing BODIPY-FL-C12 and helpful discussions. From the Puchner lab, we thank Maria-Paz Ramirez Lopez for making the Sec63 yeast strains, Ragnar Stefansson for help with data analysis. We also thank Jochen Mueller’s lab for providing the U2OS cells and Christer Ejsing for helpful discussions. Research reported in this publication was supported by the National Institute of General Medical Sciences of the National Institutes of Health under Award number R21GM127965.
Publisher Copyright:
© 2019, The Author(s).