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Conventional BODIPY Conjugates for Live-Cell Super-Resolution Microscopy and Single-Molecule Tracking
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生物化学
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JoVE 杂志 生物化学
Conventional BODIPY Conjugates for Live-Cell Super-Resolution Microscopy and Single-Molecule Tracking

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

DOI:
10.3791/60950-v

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07:49 min

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June 08, 2020

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

Chapters

  • 00:04Introduction
  • 01:24Preparation of Mammalian Cells for SMLM (Single-Molecule Localization Microscopy) Imaging
  • 02:10Equipment Preparation
  • 03:22Data Acquisition
  • 04:18Data Analysis and Single-Molecule Tracking
  • 05:31Results: SMLM Imaging Using BODIPY Conjugates
  • 06:38Conclusion

Summary

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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.

Tags

Keywords: BODIPY ConjugatesSuper-resolution MicroscopySingle-molecule TrackingLive-cell ImagingLipid DropletsFatty AcidYeast CellsMammalian CellsEMCCD CameraLaser PowerMicroscope Optimization

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