Microfluidic, Marker-free Isolation of Circulating Tumor Cells from Blood Samples

Overview

Journal Nat Protoc

Specialties Biology
Pathology
Science

Date 2014 Mar 1

PMID 24577360

Citations 267

Authors

Nezihi Murat Karabacak

Philipp S Spuhler

Fabio Fachin

Eugene J Lim

Vincent Pai

Emre Ozkumur

Joseph M Martel

Nikola Kojic

Kyle Smith

Pin-I Chen

Jennifer Yang

Henry Hwang

Bailey Morgan

Julie Trautwein

Thomas A Barber

Shannon L Stott

Shyamala Maheswaran

Ravi Kapur

Daniel A Haber

Mehmet Toner

Affiliations

Soon will be listed here.

Abstract

The ability to isolate and analyze rare circulating tumor cells (CTCs) has the potential to further our understanding of cancer metastasis and enhance the care of cancer patients. In this protocol, we describe the procedure for isolating rare CTCs from blood samples by using tumor antigen-independent microfluidic CTC-iChip technology. The CTC-iChip uses deterministic lateral displacement, inertial focusing and magnetophoresis to sort up to 10⁷ cells/s. By using two-stage magnetophoresis and depletion antibodies against leukocytes, we achieve 3.8-log depletion of white blood cells and a 97% yield of rare cells with a sample processing rate of 8 ml of whole blood/h. The CTC-iChip is compatible with standard cytopathological and RNA-based characterization methods. This protocol describes device production, assembly, blood sample preparation, system setup and the CTC isolation process. Sorting 8 ml of blood sample requires 2 h including setup time, and chip production requires 2-5 d.

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