title: High yield patterning of single cells from extremely small populations.
abstract: Many biological assays require the ability to isolate and process single cells. Some research fields, such as the characterization of rare cells, the in vitro processing of stem cells, and the study of early stage cell differentiation, call for the additional and typically unmet ability to work with extremely low-count cell populations. In all these cases, efficient single-cell handling must be matched with the ability to work on a limited number of cells with a low cell loss rate. In this paper, we present a platform combining flow-through processing with deterministic (nonstatistical) patterning of cells coming from extremely small cell populations. We describe here modules using dielectrophoresis to control the position of cells flowing in microchannels and to pattern them in open microwells where cells were further analyzed. K562 cells continuously flowing at a speed of up to 100 μm/s were tridimensionally focused, aligned, and patterned inside microwells. A high-patterning yield and low cell loss rate were demonstrated experimentally: 15uL drops, containing an average of 15 cells, were transferred to the microchannel with an 83% yield, and cells were then patterned into microwells with a 100% yield. The deterministic patterning of cells was demonstrated both by isolating single cells in microwells and by creating clusters composed of a predetermined number of cells. Cell proliferation was assessed by easily recovering cells from open microwells, and a growth rate comparable to the control was obtained.
authors: ARCES-University of Bologna, Bologna, Italy. afaenza@arces.unibo.it
authors: Andrea
authors: A
authors: Faenza
authors: Massimo
authors: M
authors: Bocchi
authors: Enri
authors: E
authors: Duqi
authors: Luca
authors: L
authors: Giulianelli
authors: Nicola
authors: N
authors: Pecorari
authors: Laura
authors: L
authors: Rambelli
authors: Roberto
authors: R
authors: Guerrieri
mesh_terms: D002454:Cell Differentiation / Q000502:physiology; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival / Q000502:physiology; D006801:Humans; D020014:K562 Cells; D046210:Microfluidic Analytical Techniques / Q000379:methods*