A rapid, gentle and scalable method for dissociation and fluorescent sorting of imaginal disc cells for mRNA sequencing

Sumbul Jawed Khan, Syeda Nayab Fatima Abidi, Yuan Tian, Andrea Skinner, Rachel K. Smith-Bolton

Research output: Contribution to journalArticle

Abstract

ABSTRACT: Dissociation of imaginal disc cells has been carried out previously to enable flow cytometry and cell sorting to analyze cell cycle progression, cell size, gene expression, and other aspects of imaginal tissues. However, the lengthy dissociation protocols employed may alter gene expression, cell behavior and overall viability. Here we describe a new rapid and gentle method of dissociating the cells of wing imaginal discs that significantly enhances cell viability and reduces the likelihood of gene expression changes. Furthermore, this method is scalable, enabling collection of large amounts of sample for high-throughput experiments without the need for data-distorting amplifications.

Original languageEnglish (US)
Pages (from-to)73-80
Number of pages8
JournalFly
Volume10
Issue number2
DOIs
StatePublished - Apr 2 2016

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Keywords

  • Dissociation
  • Drosophila
  • FACS
  • imaginal disc
  • mRNA-seq

ASJC Scopus subject areas

  • Insect Science

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