Microfluidic sorting and multimodal typing of cancer cells in self-assembled magnetic arrays.
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.
Saliba
Antoine-Emmanuel
AE
Institut Curie, Centre National de la Recherche Scientifique, Université Pierre et Marie Curie, Unité Mixte de Recherche 168, 75005 Paris, France.
Saias
Laure
L
Psychari
Eleni
E
Minc
Nicolas
N
Simon
Damien
D
Bidard
François-Clément
FC
Mathiot
Claire
C
Pierga
Jean-Yves
JY
Fraisier
Vincent
V
Salamero
Jean
J
Saada
Véronique
V
Farace
Françoise
F
Vielh
Philippe
P
Malaquin
Laurent
L
Viovy
Jean-Louis
JL
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