Biochemical and High Throughput Microscopic Assessment of Fat Mass in Caenorhabditis Elegans

Elizabeth C. Pino; Christopher M. Webster; Christopher E. Carr; Alexander A. Soukas

doi:10.3791/50180

Journal

/

Biology

/

Biokemisk og High Throughput Mikroskopisk vurdering af Fat Mass i Caenorhabditis elegans</em

JoVE Journal
Biology

This content is Free Access.

JoVE Journal Biology

Biochemical and High Throughput Microscopic Assessment of Fat Mass in Caenorhabditis Elegans

Biokemisk og High Throughput Mikroskopisk vurdering af Fat Mass i Caenorhabditis elegans</em

DOI:

10.3791/50180-v

•

16:07 min

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March 30, 2013

•

Elizabeth C. Pino, Christopher M. Webster, Christopher E. Carr, Alexander A. Soukas

¹Center for Human Genetic Research and Department of Medicine,Massachusetts General Hospital and Harvard Medical School, ²Department of Earth, Atmospheric, and Planetary Sciences,Massachusetts Institute of Technology

Chapters

00:05Title
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

Automatic Translation

Vi præsenterer robuste biokemiske og mikroskopiske metoder til undersøgelse Caenorhabditis elegans Lipid butikker. En hurtig, simpel, fastsættelse-farvningsprocedure til fluorescerende lipid dråbe imaging udnytter de spektrale egenskaber af det lipofile farvestof Nile red. Derefter præsenterer vi biokemisk måling af triglycerider og phospholipider ved hjælp af fastfase-ekstraktion og gaschromatografi-massespektrometri.

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