Conjugate

Biology

Materials science

Microscopy

Biochemistry

Molecule

Endoplasmic reticulum

Super-resolution microscopy

Physics

Nanotechnology

Organic chemistry

Mathematics

Optics

Fluorescence

Mathematical analysis

Chemistry

Fluorescence microscope

Nanoscopic scale

Biophysics

BODIPY

Nature Communications

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 (D II ), which transiently form through bi-molecular encounters and emit bright single-molecule fluorescence. We employ the versatility of D II -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.

Single-molecule localization microscopy and tracking with red-shifted states of conventional BODIPY conjugates in living cells