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

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
Pages185-193
ISBN (Electronic)9781493994120
ISBN (Print)9781493994113
DOIs
Publication statusE-pub ahead of print - 12 Apr 2019

Publication series

NameMethods in Molecular Biology
Volume1976

Cite this

Subkhankulova, T., & Kelsh, R. N. (2019). Fluorescence-activated Cell Sorting and NanoString profiling of Single Neural Crest Cells and Pigment Cells. In Q. Schwartz, & S. Wizniak (Eds.), Methods Relevant to Neural Crest Research (pp. 185-193). (Methods in Molecular Biology; Vol. 1976). Springer. https://doi.org/10.1007/978-1-4939-9412-0_14

Fluorescence-activated Cell Sorting and NanoString profiling of Single Neural Crest Cells and Pigment Cells. / Subkhankulova, Tatiana; Kelsh, Robert N.

Methods Relevant to Neural Crest Research. ed. / Quentin Schwartz; Sophie Wizniak. Springer, 2019. p. 185-193 (Methods in Molecular Biology; Vol. 1976).

Research output: Chapter in Book/Report/Conference proceedingChapter

Subkhankulova, T & Kelsh, RN 2019, Fluorescence-activated Cell Sorting and NanoString profiling of Single Neural Crest Cells and Pigment Cells. in Q Schwartz & S Wizniak (eds), Methods Relevant to Neural Crest Research. Methods in Molecular Biology, vol. 1976, Springer, pp. 185-193. https://doi.org/10.1007/978-1-4939-9412-0_14
Subkhankulova T, Kelsh RN. Fluorescence-activated Cell Sorting and NanoString profiling of Single Neural Crest Cells and Pigment Cells. In Schwartz Q, Wizniak S, editors, Methods Relevant to Neural Crest Research. Springer. 2019. p. 185-193. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-9412-0_14
Subkhankulova, Tatiana ; Kelsh, Robert N. / Fluorescence-activated Cell Sorting and NanoString profiling of Single Neural Crest Cells and Pigment Cells. Methods Relevant to Neural Crest Research. editor / Quentin Schwartz ; Sophie Wizniak. Springer, 2019. pp. 185-193 (Methods in Molecular Biology).
@inbook{7bef00953e6245068dc5538fa4b4db54,
title = "Fluorescence-activated Cell Sorting and NanoString profiling of Single Neural Crest Cells and Pigment Cells",
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.",
author = "Tatiana Subkhankulova and Kelsh, {Robert N.}",
year = "2019",
month = "4",
day = "12",
doi = "10.1007/978-1-4939-9412-0_14",
language = "English",
isbn = "9781493994113",
series = "Methods in Molecular Biology",
publisher = "Springer",
pages = "185--193",
editor = "Quentin Schwartz and Sophie Wizniak",
booktitle = "Methods Relevant to Neural Crest Research",

}

TY - CHAP

T1 - Fluorescence-activated Cell Sorting and NanoString profiling of Single Neural Crest Cells and Pigment Cells

AU - Subkhankulova, Tatiana

AU - Kelsh, Robert N.

PY - 2019/4/12

Y1 - 2019/4/12

N2 - 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.

AB - 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.

U2 - 10.1007/978-1-4939-9412-0_14

DO - 10.1007/978-1-4939-9412-0_14

M3 - Chapter

SN - 9781493994113

T3 - Methods in Molecular Biology

SP - 185

EP - 193

BT - Methods Relevant to Neural Crest Research

A2 - Schwartz, Quentin

A2 - Wizniak, Sophie

PB - Springer

ER -