Microscale Laminar Vortices for High-Purity Extraction and Release of Circulating Tumor Cells

Methods Mol Biol. 2017:1634:65-79. doi: 10.1007/978-1-4939-7144-2_5.

Abstract

Circulating tumor cells (CTCs) are disseminated tumor cells that reflect the tumors of origin and can provide a liquid biopsy that would potentially enable noninvasive tumor profiling, treatment monitoring, and identification of targeted treatments. Accurate and rapid purification of CTCs holds great potential to improve cancer care but the task remains technically challenging. Microfluidic isolation of CTCs within microscale vortices enables high-throughput and size-based purification of rare CTCs from bodily fluids. Collected cells are highly pure, viable, and easily accessible, allowing seamless integration with various downstream applications. Here, we describe how to fabricate the High-Throughput Vortex Chip (Vortex-HT) and to process diluted whole blood for CTC collection. Lastly, immunostaining and imaging protocols for CTC classification and corresponding CTC image galleries are reported.

Keywords: CTC enumeration; CTC purification; High-throughput size-based cell separation; Microscale laminar vortices.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / metabolism
  • Biomarkers, Tumor / genetics
  • Biomarkers, Tumor / immunology
  • Biomarkers, Tumor / metabolism
  • Cell Count
  • Cell Separation / instrumentation
  • Cell Separation / methods*
  • Cell Size
  • Dimethylpolysiloxanes / chemistry
  • Equipment Design*
  • Fluorescein-5-isothiocyanate / chemistry
  • Fluorescent Antibody Technique / methods
  • Fluorescent Dyes / chemistry
  • Humans
  • Immunoconjugates / chemistry
  • Keratins / genetics
  • Keratins / immunology
  • Keratins / metabolism
  • Lab-On-A-Chip Devices*
  • Leukocyte Common Antigens / genetics
  • Leukocyte Common Antigens / immunology
  • Leukocyte Common Antigens / metabolism
  • Microfluidic Analytical Techniques / instrumentation*
  • Neoplasms / blood
  • Neoplasms / diagnosis*
  • Neoplasms / immunology
  • Neoplasms / pathology
  • Neoplastic Cells, Circulating / immunology
  • Neoplastic Cells, Circulating / metabolism
  • Neoplastic Cells, Circulating / pathology*
  • Nylons / chemistry
  • Phycoerythrin / chemistry
  • Protein Binding
  • Rheology

Substances

  • Antibodies, Monoclonal
  • Biomarkers, Tumor
  • Dimethylpolysiloxanes
  • Fluorescent Dyes
  • Immunoconjugates
  • Nylons
  • poly(dimethylsiloxane)-polyamide copolymer
  • Phycoerythrin
  • Keratins
  • Leukocyte Common Antigens
  • PTPRC protein, human
  • Fluorescein-5-isothiocyanate