Single Cell Manipulation Technology-Towards "Ink-Jet" Printing of Cancer Arrays
Victoria McEneaney, Helen Keegan, Michael Gallagher, Cara Martin, Orla Sheils, Jonas Schondube, Andre Gross, Peter Koltay, John O'Leary. Trinity College Dublin, Dublin, Leinster, Ireland; IMTEK, Freiburg, Germany
Background: An ongoing EU Framework 7 project (PASCA: Platform for Advanced Single Cell Manipulation and Analysis) has developed a novel instrument referred to as a Single Cell Manipulator (SCM) device which enables 1) isolation of single cells from a cell suspension 2) generation of a picoliter sized droplet containing the single cell and 3) printing of the single cell in an “ink-jet” like manner in an automated, ordered array onto a chosen substrate for subsequent downstream analysis.
Design: The biological or clinical samples are injected into the silicon dispenser chip, coupled to a live cell camera that images and displays cells approaching the chip exit nozzle. An optical particle detection mechanism determines the presence of single cells within a selected region of interest, whilst a sorting algorithm ensures that only droplets containing single cells are selected for printing to the prescribed location. The SCM will be further developed within the project to allow sorting of cells based on their electrical impedance properties or with the detection of fluorescently labelled tags.
Results: Applications currently undergoing analysis on SCM technology include - but are not exclusive to – cervical cytopathology, anaplastic thyroid carcinoma and cancer stem cells. In cervical cytopathology, single HeLa cells have been printed and successfully cultured in 96 well plates. Separation of HPV negative and HPV positive cervical cells from mixed populations with subsequent printing of the isolated cells into ordered arrays is ongoing, with a view to progressing to printing arrays of clinical cytology samples onto microscopy slides. Single WT thyrocytes and anaplastic thyroid carcinoma cell lines have been successfully printed and cultured in 96 well plates. Separations of WT thyrocytes and anaplastic thyroid carcinoma cells have been performed from mixed populations on the SCM based on size properties and a 100% cell selection success rate has been confirmed with fluorescence microscopy.
Conclusions: The SCM PASCA technology allows isolation of single cells from a heterogeneous population followed by ordered automated printing of these cells onto a chosen substrate. The device is further being developed to allow the printing of cancer arrays whereby the cells in a clinical sample are spatially arranged according to a gradient of abnormality. This technology will assist in analysis of individual cells within many tumours and disease types, thus leading to a greater understanding of the complexity within clinical diagnostics and therapeutics.
Tuesday, March 5, 2013 1:30 PM
Proffered Papers: Section E, Tuesday Afternoon