Open AccessCombining Laser Capture Microdissection and Microfluidic qPCR to Analyze Transcriptional Profiles of Single Cells: A Systems Biology Approach to Opioid Dependence
Author(s) -
Sean O'Sullivan,
Beverly A.S. Reyes,
Rajanikanth Vadigepalli,
Elisabeth J. Van Bockstaele,
James S. Schwaber
Publication year - 2020
Publication title -
journal of visualized experiments
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.596
H-Index - 91
ISSN - 1940-087X
DOI - 10.3791/60612
Subject(s) - laser capture microdissection , biology , single cell analysis , microfluidics , computational biology , microdissection , gene expression , phenotype , microbiology and biotechnology , cell , gene , genetics , nanotechnology , materials science
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.