Preparation of Mammalian Cells for SMLM (Single-Molecule Localization Microscopy) Imaging
2:10
Equipment Preparation
3:22
Data Acquisition
4:18
Data Analysis and Single-Molecule Tracking
5:31
Results: SMLM Imaging Using BODIPY Conjugates
6:38
Conclusion
Trascrizione
Our protocol describes the use of conventional BODIPY dyes for super-resolution microscopy using their sparse, red-shifted states. This enables us to study organelles and biomolecules in living cells with 30-nanometer resolution. The advantage of
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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.