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Biochemical and High Throughput Microscopic Assessment of Fat Mass in Caenorhabditis Elegans
Journal JoVE
Biologie
This content is Free Access.
Journal JoVE Biologie
Biochemical and High Throughput Microscopic Assessment of Fat Mass in Caenorhabditis Elegans
DOI:

16:07 min

March 30, 2013

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Chapitres

  • 00:05Titre
  • 02:06Stamp Frozen Library Plates onto Rectangular Amp/Tet LB Agar Plates
  • 03:16Grow and Seed RNAi Bacterial Clones and Add Worms to RNAi Plates
  • 05:49Fix and Stain Worms in Nile Red and Image on a High-throughput Scope
  • 07:22Wash Worms and Image on a High-throughput Microscope
  • 08:31Biochemical Lipid Determination Using Solid Phase Extraction and Gas Chromatography-mass Spectrometry
  • 12:02Representative Fat Mass Content Results
  • 15:39Conclusion

Summary

Traduction automatique

We present robust biochemical and microscopic methods for studying Caenorhabditis elegans lipid stores. A rapid, simple, fixing-staining procedure for fluorescent lipid droplet imaging leverages the spectral properties of the lipophilic dye Nile red. We then present biochemical measurement of triglycerides and phospholipids using solid phase extraction and gas chromatography-mass spectrometry.

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