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Peer-reviewed veterinary case report

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

Journal:
Journal of visualized experiments : JoVE
Year:
2020
Authors:
O'Sullivan, Sean J et al.
Affiliation:
Daniel Baugh Institute for Functional Genomics and Computational Biology

Abstract

Profound transcriptional heterogeneity in anatomically adjacent single cells suggests that robust tissue functionality may be achieved by cellular phenotype diversity. Single-cell experiments investigating the network dynamics of biological systems demonstrate cellular and tissue responses to various conditions at biologically meaningful resolution. Herein, we explain our methods for gathering single cells from anatomically specific locations and accurately measuring a subset of their gene expression profiles. We combine laser capture microdissection (LCM) with microfluidic reverse transcription quantitative polymerase chain reactions (RT-qPCR). We also use this microfluidic RT-qPCR platform to measure the microbial abundance of gut contents.

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Original publication: https://pubmed.ncbi.nlm.nih.gov/32202523/