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University of Minnesota, Twin Cities, Physics and Nanotechnology (PAN)

2 ARTICLES PUBLISHED IN JoVE

Biochemistry

Conventional BODIPY Conjugates for Live-Cell Super-Resolution Microscopy and Single-Molecule Tracking

Santosh Adhikari¹, Chiranjib Banerjee^*1, Joe Moscatelli^*1,2, Elias M. Puchner¹

¹School of Physics and Astronomy, University of Minnesota, Twin Cities, Physics and Nanotechnology (PAN), ²Middlebury College

Conventional BODIPY conjugates can be used for live-cell single-molecule localization microscopy (SMLM) through exploitation of their transiently forming, red-shifted ground state dimers. We present an optimized SMLM protocol to track and resolve subcellular neutral lipids and fatty acids in living mammalian and yeast cells at the nanoscopic length scale.

Biochemistry

Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET

Chiranjib Banerjee¹, Brandon Wey-Hung Liauw¹, Reza Vafabakhsh¹

¹Department of Molecular Biosciences, Northwestern University

This study presents a detailed procedure to perform single-molecule fluorescence resonance energy transfer (smFRET) experiments on G protein-coupled receptors (GPCRs) using site-specific labeling via unnatural amino acid (UAA) incorporation. The protocol provides a step-by-step guide for smFRET sample preparation, experiments, and data analysis.