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JoVE Journal Genetics
Combining Laser Capture Microdissection and Microfluidic qPCR to Analyze Transcriptional Profiles of Single Cells
 

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

Article DOI: 10.3791/60612-v • 09:54 min • March 8th, 2020
March 8th, 2020 •
1,2, 3, 1, 3, 1
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

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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.

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Laser Capture Microdissection Microfluidic QPCR Transcriptional Profiles Single Cells Systems Biology Approach Opioid Dependence Gene Expression Cellular Subphenotypes Anatomic Specificity Biological Problem Individual Cellular Response Neurons Cell Types Disease Disturbance Function Brain Region Cryostat Chuck Amygdala Tissue Collection Box Glass Slides Metal Pan Ethanol Dehydration Series
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