title: Microfluidic, marker-free isolation of circulating tumor cells from blood samples.
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.
authors.affiliation: 1] Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA. [2].
authors.forename: Nezihi Murat
authors.initials: NM
authors.lastname: Karabacak
authors.affiliation: 1] Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA. [2].
authors.forename: Philipp S
authors.initials: PS
authors.lastname: Spuhler
authors.affiliation: Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Fabio
authors.initials: F
authors.lastname: Fachin
authors.affiliation: Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Eugene J
authors.initials: EJ
authors.lastname: Lim
authors.affiliation: Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Vincent
authors.initials: V
authors.lastname: Pai
authors.affiliation: Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Emre
authors.initials: E
authors.lastname: Ozkumur
authors.affiliation: Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Martel
authors.affiliation: Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Nikola
authors.initials: N
authors.lastname: Kojic
authors.affiliation: Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Kyle
authors.initials: K
authors.lastname: Smith
authors.affiliation: Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Pin-i
authors.initials: PI
authors.lastname: Chen
authors.affiliation: Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Henry
authors.initials: H
authors.lastname: Hwang
authors.affiliation: Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Bailey
authors.initials: B
authors.lastname: Morgan
authors.affiliation: Cancer Center, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Julie
authors.initials: J
authors.lastname: Trautwein
authors.affiliation: Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Barber
authors.affiliation: 1] Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA. [2] Cancer Center, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Shannon L
authors.initials: SL
authors.lastname: Stott
authors.affiliation: Cancer Center, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Shyamala
authors.initials: S
authors.lastname: Maheswaran
authors.affiliation: Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Ravi
authors.initials: R
authors.lastname: Kapur
authors.affiliation: 1] Cancer Center, Massachusetts General Hospital, Boston, Massachusetts, USA. [2] Howard Hughes Medical Institute, Chevy Chase, Maryland, USA.
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Haber
authors.affiliation: Department of Surgery and Center for Engineering in Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Mehmet
authors.initials: M
authors.lastname: Toner
mesh_terms: D002469:Cell Separation / Q000379:methods*; D006801:Humans; D019476:Insect Proteins; D059346:Magnets; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D009360:Neoplastic Cells, Circulating
chemical_list: C116357:ICHIT protein, Anopheles gambiae; D019476:Insect Proteins