Journal
/
遗传学
/
Combining Laser Capture Microdissection and Microfluidic qPCR to Analyze Transcriptional Profiles of Single Cells: A Systems Biology Approach to Opioid Dependence
JoVE 杂志
遗传学
需要订阅 JoVE 才能查看此.  登录或开始免费试用。
JoVE 杂志 遗传学
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

•

09:54 min

•

March 08, 2020

•

, , , ,
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

自动翻译

自动翻译

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

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

Article

Embed

添加到播放列表

Usage Statistics

Related Videos

Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards

Single Molecule Analysis of Laser Localized Psoralen Adducts

Laser Microirradiation to Study In Vivo Cellular Responses to Simple and Complex DNA Damage

Adaptation of Hybridization Capture of Chromatin-associated Proteins for Proteomics to Mammalian Cells

Ultralow Input Genome Sequencing Library Preparation from a Single Tardigrade Specimen

A Novel Saturation Mutagenesis Approach: Single Step Characterization of Regulatory Protein Binding Sites in RNA Using Phosphorothioates

The Replica Set Method: A High-throughput Approach to Quantitatively Measure Caenorhabditis elegans Lifespan

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

Generation of Cancer Cell Clones to Visualize Telomeric Repeat-containing RNA TERRA Expressed from a Single Telomere in Living Cells

Read Article