Microfluidic, marker-free isolation of circulating tumor cells from blood samples
| Autor: | Nikola Kojic, Kyle C. Smith, Shyamala Maheswaran, Vincent Pai, Bailey Morgan, Henry H. Hwang, Julie Trautwein, Fabio Fachin, Emre Ozkumur, Joseph M. Martel, Pin-i Chen, Philipp S. Spuhler, Shannon L. Stott, Tom Barber, Jennifer Yang, Ravi Kapur, Mehmet Toner, Eugene J. Lim, Daniel A. Haber, Nezihi Murat Karabacak |
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| Rok vydání: | 2014 |
| Předmět: |
Microfluidics
Cancer Cell Separation Microfluidic Analytical Techniques Biology Neoplastic Cells Circulating Isolation (microbiology) medicine.disease Molecular biology Article General Biochemistry Genetics and Molecular Biology Circulating tumor cell Magnets medicine biology.protein Humans Insect Proteins Sample preparation Marker free Antibody Whole blood |
| Zdroj: | Nature Protocols. 9:694-710 |
| ISSN: | 1750-2799 1754-2189 |
| DOI: | 10.1038/nprot.2014.044 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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