High-Throughput Isolation of Cell Protrusions with Single-Cell Precision for Profiling Subcellular Gene Expression

Pengchao Zhang, Xin Han, Jun Yao, Ning Shao, Kai Zhang, Yufu Zhou, Youli Zu, Bin Wang, Lidong Qin

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Invading cancer cells extend cell protrusions, which guide cancer-cell migration and invasion, eventually leading to metastasis. The formation and activity of cell protrusions involve the localization of molecules and organelles at the cell front; however, it is challenging to precisely isolate these subcellular structures at the single-cell level for molecular analysis. Here, we describe a newly developed microfluidic platform capable of high-throughput isolation of cell protrusions at single-cell precision for profiling subcellular gene expression. Using this microfluidic platform, we demonstrate the efficient generation of uniform cell-protrusion arrays (more than 5000 cells with protrusions) for a series of cell types. We show precise isolation of cell protrusions with high purity at single-cell precision for subsequent RNA-Seq analysis, which was further validated by RT-qPCR and RNA FISH. Our highly controlled protrusion isolation method opens a new avenue for the study of subcellular functional mechanisms and signaling pathways in metastasis.

Original languageEnglish (US)
Pages (from-to)13700-13705
Number of pages6
JournalAngewandte Chemie - International Edition
Volume58
Issue number39
DOIs
StatePublished - Sep 23 2019

Keywords

  • RNA sequencing
  • analytical methods
  • cell protrusion
  • gene expression
  • microfluidics

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)

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