title: Microfluidic sorting and multimodal typing of cancer cells in self-assembled magnetic arrays.
abstract: We propose a unique method for cell sorting, "Ephesia," using columns of biofunctionalized superparamagnetic beads self-assembled in a microfluidic channel onto an array of magnetic traps prepared by microcontact printing. It combines the advantages of microfluidic cell sorting, notably the application of a well controlled, flow-activated interaction between cells and beads, and those of immunomagnetic sorting, notably the use of batch-prepared, well characterized antibody-bearing beads. On cell lines mixtures, we demonstrated a capture yield better than 94%, and the possibility to cultivate in situ the captured cells. A second series of experiments involved clinical samples--blood, pleural effusion, and fine needle aspirates--issued from healthy donors and patients with B-cell hematological malignant tumors (leukemia and lymphoma). The immunophenotype and morphology of B-lymphocytes were analyzed directly in the microfluidic chamber, and compared with conventional flow cytometry and visual cytology data, in a blind test. Immunophenotyping results using Ephesia were fully consistent with those obtained by flow cytometry. We obtained in situ high resolution confocal three-dimensional images of the cell nuclei, showing intranuclear details consistent with conventional cytological staining. Ephesia thus provides a powerful approach to cell capture and typing allowing fully automated high resolution and quantitative immunophenotyping and morphological analysis. It requires at least 10 times smaller sample volume and cell numbers than cytometry, potentially increasing the range of indications and the success rate of microbiopsy-based diagnosis, and reducing analysis time and cost.
authors.affiliation: Institut Curie, Centre National de la Recherche Scientifique, Université Pierre et Marie Curie, Unité Mixte de Recherche 168, 75005 Paris, France.
authors.forename: Antoine-Emmanuel
authors.initials: AE
authors.lastname: Saliba
authors.forename: Laure
authors.initials: L
authors.lastname: Saias
authors.forename: Eleni
authors.initials: E
authors.lastname: Psychari
authors.forename: Nicolas
authors.initials: N
authors.lastname: Minc
authors.forename: Damien
authors.initials: D
authors.lastname: Simon
authors.forename: François-Clément
authors.initials: FC
authors.lastname: Bidard
authors.forename: Claire
authors.initials: C
authors.lastname: Mathiot
authors.forename: Jean-Yves
authors.initials: JY
authors.lastname: Pierga
authors.forename: Vincent
authors.initials: V
authors.lastname: Fraisier
authors.forename: Jean
authors.initials: J
authors.lastname: Salamero
authors.forename: Véronique
authors.initials: V
authors.lastname: Saada
authors.forename: Françoise
authors.initials: F
authors.lastname: Farace
authors.forename: Philippe
authors.initials: P
authors.lastname: Vielh
authors.forename: Laurent
authors.initials: L
authors.lastname: Malaquin
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Viovy
mesh_terms: D000465:Algorithms; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D005434:Flow Cytometry; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D016130:Immunophenotyping; D019169:Jurkat Cells; D008280:Magnetics; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D018625:Microscopy, Atomic Force; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D008962:Models, Theoretical; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology