title: A rapid, gentle and scalable method for dissociation and fluorescent sorting of imaginal disc cells for mRNA sequencing.
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.
authors.affiliation: a Department of Cell and Developmental Biology , University of Illinois at Urbana-Champaign , Urbana , IL , USA.
authors.forename: Sumbul Jawed
authors.initials: SJ
authors.lastname: Khan
authors.affiliation: a Department of Cell and Developmental Biology , University of Illinois at Urbana-Champaign , Urbana , IL , USA.
authors.forename: Syeda Nayab Fatima
authors.initials: SN
authors.lastname: Abidi
authors.affiliation: a Department of Cell and Developmental Biology , University of Illinois at Urbana-Champaign , Urbana , IL , USA.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Tian
authors.affiliation: a Department of Cell and Developmental Biology , University of Illinois at Urbana-Champaign , Urbana , IL , USA.
authors.forename: Andrea
authors.initials: A
authors.lastname: Skinner
authors.affiliation: a Department of Cell and Developmental Biology , University of Illinois at Urbana-Champaign , Urbana , IL , USA.
authors.forename: Rachel K
authors.initials: RK
authors.lastname: Smith-Bolton
mesh_terms: D000818:Animals; D005434:Flow Cytometry / Q000379:methods*; D020869:Gene Expression Profiling; D060227:Imaginal Discs / Q000166:cytology*; D012333:RNA, Messenger / Q000235:genetics; D017423:Sequence Analysis, RNA
chemical_list: D012333:RNA, Messenger
keywords: Dissociation; Drosophila; FACS; imaginal disc; mRNA-seq