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Combining Laser Capture Microdissection and Microfluidic qPCR to Analyze Transcriptional Profiles of Single Cells: A Systems Biology Approach to Opioid Dependence
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Genetics
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JoVE Journal Genetics
Combining Laser Capture Microdissection and Microfluidic qPCR to Analyze Transcriptional Profiles of Single Cells: A Systems Biology Approach to Opioid Dependence

Combining Laser Capture Microdissection and Microfluidic qPCR to Analyze Transcriptional Profiles of Single Cells: A Systems Biology Approach to Opioid Dependence

DOI:
10.3791/60612-v

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09:54 min

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March 08, 2020

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, , , ,
1Daniel Baugh Institute for Functional Genomics and Computational Biology, Department of Pathology, Anatomy, and Cell Biology,Thomas Jefferson University, 2Sidney Kimmel Medical College,Thomas Jefferson University, 3Department of Pharmacology & Physiology,Drexel University College of Medicine

Chapters

  • 00:05Introduction
  • 00:59Preparing the Samples
  • 02:16Laser Capture Microdissection
  • 04:07Single-cell Microfluidic RT-qPCR
  • 08:08Results: Microfluidic qPCR Yields Accurate and Anatomically Specific Transcriptional Profiles of Single-Cells
  • 09:21Conclusion

Summary

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This protocol explains how to collect single neurons, microglia, and astrocytes from the central nucleus of the amygdala with high accuracy and anatomic specificity using laser capture microdissection. Additionally, we explain our use of microfluidic RT-qPCR to measure a subset of the transcriptome of these cells.

Tags

Keywords: Laser Capture MicrodissectionMicrofluidic QPCRTranscriptional ProfilesSingle CellsSystems BiologyOpioid DependenceGene ExpressionCellular SubphenotypesAnatomic SpecificityBiological ProblemsNeuronsBrain RegionsAmygdalaEthanol DehydrationXyleneFluorescenceLCM SoftwareInfrared Laser

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