Fluorescence-activated Cell Sorting and NanoString profiling of Single Neural Crest Cells and Pigment Cells

Tatiana Subkhankulova, Robert N. Kelsh

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

Abstract

Neural crest cells are an important class of multipotent stem cells, generating highly diverse derivatives. Understanding the gene regulatory networks underlying this process is of great interest, but the highly migratory and thus widely dispersed nature of the differentiating cells makes isolation of cells difficult. Fluorescence-activated cell sorting (FACS) of transgenically labelled neural crest-derived cells after disaggregation of embryos is well-suited to purifying these cells. However, their diverse differentiation means that transcriptional analysis at single cell resolution is necessary to dissect the gene regulatory networks at play. NanoString technology provides a method for highly sensitive, quantitative transcriptional profiling for a pre-defined set of genes of interest. Here we provide a detailed protocol for FACS purification of neural crest-derived cells, sorted as single cells into a multi-well plate, and their subsequent NanoString profiling, using a predetermined gene set focused on pigment cells.
Original languageEnglish
Title of host publicationMethods Relevant to Neural Crest Research
EditorsQuentin Schwartz, Sophie Wizniak
PublisherSpringer
Chapter14
Pages185-193
Number of pages9
Volume1976
ISBN (Electronic)9781493994120
ISBN (Print)9781493994113
DOIs
Publication statusPublished - 2019

Publication series

NameMethods in Molecular Biology
Volume1976

Keywords

  • NanoString
  • Neural crest
  • Pigment cells
  • Single cell expression
  • Zebrafish
  • nCounter

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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