期刊
NATURE PROTOCOLS
卷 9, 期 3, 页码 694-710出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2014.044
关键词
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资金
- US National Institutes of Health (NIH) P41 Resource Center
- NIH National Institute of Biomedical Imaging and Bioengineering Quantum Grant
- Stand Up to Cancer
- Howard Hughes Medical Institute
- Prostate Cancer Foundation and the Charles Evans Foundation
- Johnson and Johnson
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.
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