Microfluidic, marker-free isolation of circulating tumor cells from blood samples

• Published in: Nature protocols Vol. 9; no. 3; pp. 694 - 710
• Main Authors: Karabacak, Nezihi Murat, Spuhler, Philipp S, Fachin, Fabio, Lim, Eugene J, Pai, Vincent, Ozkumur, Emre, Martel, Joseph M, Kojic, Nikola, Smith, Kyle, Chen, Pin-i, Yang, Jennifer, Hwang, Henry, Morgan, Bailey, Trautwein, Julie, Barber, Thomas A, Stott, Shannon L, Maheswaran, Shyamala, Kapur, Ravi, Haber, Daniel A, Toner, Mehmet
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.03.2014; Nature Publishing Group
• Subjects: 631/1647/2230; 631/1647/350/877; 631/67/2322; Analysis; Analytical Chemistry; Antibodies; Biological Techniques; Blood; Cancer; Cell Separation - methods; Cells; Computational Biology/Bioinformatics; Humans; Insect Proteins; Leukocytes; Life Sciences; Magnets; Metastasis; Microarrays; Microfluidic Analytical Techniques - methods; Neoplastic Cells, Circulating; Organic Chemistry; Physiological aspects; protocol; Sample preparation; Tumor antigens; Tumors; China; United States > US; Massachusetts
• ISSN: 1754-2189; 1750-2799
• DOI: 10.1038/nprot.2014.044

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 10 7 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.