Journal: British Journal of Cancer

Article Title: Semiautomated isolation and molecular characterisation of single or highly purified tumour cells from CellSearch enriched blood samples using dielectrophoretic cell sorting

doi: 10.1038/bjc.2013.92

Figure Lengend Snippet: Schematic of overall experimental workflow. Step 1: Tumour cells of three different human cultured breast cancer cell lines were spiked in healthy donor blood at a concentration of 10 3 tumour cells per 7.5 ml whole blood. Step 2: Tumour cells were immunomagnetically enriched using either the CellSearch CTC kit or the CellSearch Profile kit followed by a manual staining procedure. Step 3: Cells were reconstituted in a final volume of 14 μ l and loaded in a DEPArray cartridge. Step 4: Analysis and sorting procedures were performed on the DEPArray system. Step 5: Single cells and groups of cells of interest were isolated with the DEPArray system. Step 6: Mutation or transcriptional analysis of isolated tumour cells.

Article Snippet: The sample was processed with the CellSearch CTC kit and the CellSearch cartridge was stored at 4 °C for 8 days.

Techniques: Cell Culture, Concentration Assay, Staining, Isolation, Mutagenesis

Journal: British Journal of Cancer

Article Title: Semiautomated isolation and molecular characterisation of single or highly purified tumour cells from CellSearch enriched blood samples using dielectrophoretic cell sorting

doi: 10.1038/bjc.2013.92

Figure Lengend Snippet: Composite image gallery showing staining characteristics of MDA-MB-231 cells and WBCs from a CellSearch cartridge after immunomagnetic enrichment with the CellSearch CTC kit visualised on the DEPArray system. ( A ) Tumour cells were defined using standard CellSearch CTC criteria being (1) round to oval shape, (2) presence of a clear DAPI-stained nucleus, (3) at least 50% overlap between the CK-PE-positive cytoplasm and the nucleus and (4) CD45-APC negativity. Separate images for PE, DAPI and APC fluorescence and bright field channels and merged CK-PE/DAPI and CD45-APC/DAPI images of three single MDA-MB-231 cells (top 3 rows) and two WBC (bottom 2 rows) are shown. ( B ) Scatter plot of mean fluorescence intensities for CK-PE (y axis) and CD45-APC (x axis) staining by the DEPArray software. Rectangular gates can be applied to aid quick identification of cells or populations of cells for cell sorting. Green dots represent CK+/CD45− MDA-MB-231 cells selected by the user. Red dots represent CK−/CD45+ WBCs. Grey dots represent unselected events.

Article Snippet: The sample was processed with the CellSearch CTC kit and the CellSearch cartridge was stored at 4 °C for 8 days.

Techniques: Staining, Fluorescence, Software, FACS