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Preparation of Human Tissues Embedded in Optimal Cutting Temperature Compound for Mass Spectrometry Analysis
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Cancer Research
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JoVE Journal Cancer Research
Preparation of Human Tissues Embedded in Optimal Cutting Temperature Compound for Mass Spectrometry Analysis

Preparation of Human Tissues Embedded in Optimal Cutting Temperature Compound for Mass Spectrometry Analysis

DOI:
10.3791/62552-v

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09:09 min

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April 27, 2021

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, ,
1Department of Biology,Virginia Commonwealth University, 2VCU Lipidomics/Metabolomics Shared Resource,Virginia Commonwealth University School of Medicine, 3Massey Cancer Center

Chapters

  • 00:04Introduction
  • 00:54Preparation of Tissue-Drying Wicks
  • 01:39Tissue Washing for Sphingolipidomic Optimal Cutting Temperature-Compound (OCT) Removal
  • 02:41Tissue Weighing for Data Normalization
  • 04:12Lipid Extraction
  • 07:03Results: Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry (LC ESI MS/MS) Analysis of Tissues Following OCT Removal
  • 08:36Conclusion

Summary

Automatic Translation

Automatic Translation

Sphingolipids are bioactive metabolites with well-established roles in human disease. Characterizing alterations in tissues with mass-spectrometry can reveal roles in disease etiology or identify therapeutic targets. However, the OCT-compound used for cryopreservation in biorepositories interferes with mass-spectrometry. We outline methods to analyze sphingolipids in human tissues embedded in OCT with LC-ESI-MS/MS.

Tags

OCT Compound RemovalMass Spectrometry AnalysisSphingolipid QuantificationLiquid Chromatography-mass SpectrometryLipid Extraction

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