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Open Access Highly Accessed Methodology article

Single cell cytometry of protein function in RNAi treated cells and in native populations

Peter LaPan1, Jing Zhang2, Jing Pan2, Andrew Hill3 and Steven A Haney24*

Author Affiliations

1 Department of Biological Technologies, Section of Biologic Research, Wyeth Research, 87 Cambridge Park Drive, Cambridge, MA 02140, USA

2 Department of Biological Technologies, Oncology Research, Wyeth Research, 87 Cambridge Park Drive, Cambridge, MA 02140, USA

3 Department of Biological Technologies, Bioinformatics, Wyeth Research, 87 Cambridge Park Drive, Cambridge, MA 02140, USA

4 Pfizer Research Technology Center, 620 Memorial Drive, Cambridge, MA 02139

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BMC Cell Biology 2008, 9:43  doi:10.1186/1471-2121-9-43

Published: 1 August 2008

Additional files

Additional file 1:

Supplementary Figure 1 Effect of etoposide treatment on nuclear area of SW 480 colon carcinoma cells. SW480 cells were plated in a 96-well microtiter plate and cultured for 24 hr, at which time they were treated with 5 mM etoposide (shown in red) or a vehicle control (shown in blue). Nuclear size for each cell is shown as a histogram of the entire dataset after binning as shown in the figure.

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Additional file 2:

Immunofluorescence and siRNA transfection conditions. Specific catalog and treatment conditions for siRNAs, transfection reagents and immunofluorescence microscopy.

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This file can be viewed with: Microsoft Excel Viewer

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Additional file 3:

Supplementary Figure 2 Representative specific and non-specific staining intensities for the primary and secondary antibodies. Box plots of antigen levels as detected by high content screening. Specific antigens were detected using antibodies as indicated in the panel and non-specific background staining was detected using specific isotypes, is indicated as well. IgG is from rabbit, IgG1 and IgG2a are from mouse.

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Additional file 4:

Extracting high content single cell data for analysis. SWEAVE document describing the routine used to retrieve single cell data from the database.

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