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Nat Protoc. 2016 Jan;11(1):134-48. doi: 10.1038/nprot.2016.003. Epub 2015 Dec 17.

Ultra-fast, label-free isolation of circulating tumor cells from blood using spiral microfluidics.

Warkiani ME1,2,3, Khoo BL3,4, Wu L5, Tay AK3,6, Bhagat AA7, Han J3,5,8, Lim CT3,4,6.

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School of Mechanical and Manufacturing Engineering, Australian Center for NanoMedicine, University of New South Wales, Sydney, New South Wales, Australia.
Medical Oncology Group, Ingham Institute for Applied Medical Research, Liverpool, New South Wales, Australia.
BioSystems and Micromechanics (BioSyM), Singapore-MIT Alliance for Research and Technology (SMART) Centre, Singapore, Singapore.
Mechanobiology Institute, National University of Singapore, Singapore, Singapore.
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Department of Biomedical Engineering, National University of Singapore, Singapore, Singapore.
Clearbridge BioMedics Pte Ltd, Singapore, Singapore.
Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.


Circulating tumor cells (CTCs) are rare cancer cells that are shed from primary or metastatic tumors into the peripheral blood circulation. Phenotypic and genetic characterization of these rare cells can provide important information to guide cancer staging and treatment, and thus further research into their characteristics and properties is an area of considerable interest. In this protocol, we describe detailed procedures for the production and use of a label-free spiral microfluidic device to allow size-based isolation of viable CTCs using hydrodynamic forces that are present in curvilinear microchannels. This spiral system enables us to achieve ≥ 85% recovery of spiked cells across multiple cancer cell lines and 99.99% depletion of white blood cells in whole blood. The described spiral microfluidic devices can be produced at an extremely low cost using standard microfabrication and soft lithography techniques (2-3 d), and they can be operated using two syringe pumps for lysed blood samples (7.5 ml in 12.5 min for a three-layered multiplexed chip). The fast processing time and the ability to collect CTCs from a large patient blood volume allows this technique to be used experimentally in a broad range of potential genomic and transcriptomic applications.

[Indexed for MEDLINE]

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