Title :
Time-lapse microscopy-based genome wide RNAi screening in live human cells
Author :
Neumann, Beate ; Held, Michael ; Liebel, Urban ; Erfle, Holger ; Rogers, Phill ; Pepperkok, Rainer ; Ellenberg, Jan
Author_Institution :
MitoCheck Project Group, EMBL, Heidelberg
Abstract :
Specific knock-down of gene expression by RNA interference is the method of choice to study gene function in human cells. A uniquely detailed phenotypic readout of such hypomorphs is possible by live cell microscopy of appropriate fluorescent reporter proteins. Here, I will present a fully automated method for RNAi screens in cultured human cells, combining reverse transfection by siRNA cell arrays, automated time-lapse fluorescence microscopy and computational phenotype analysis by image processing. The strategy is illustrated using an automatically scored mitosis assay and provides an easily scalable platform that we currently use in genome-wide RNAi screens for several cellular functions
Keywords :
biological techniques; biology computing; cellular biophysics; fluorescence; genetics; image processing; molecular biophysics; optical microscopy; proteins; RNA interference; automated time-lapse fluorescence microscopy; automatically scored mitosis assay; cellular functions; computational phenotype analysis; fluorescent reporter proteins; gene expression; gene function; genome wide RNAi screening; hypomorphs; image processing; live human cells; reverse transfection; siRNA cell arrays; Bioinformatics; Cells (biology); Fluorescence; Gene expression; Genomics; Humans; Interference; Microscopy; Proteins; RNA;
Conference_Titel :
Biomedical Imaging: Nano to Macro, 2006. 3rd IEEE International Symposium on
Conference_Location :
Arlington, VA
Print_ISBN :
0-7803-9576-X
DOI :
10.1109/ISBI.2006.1624898
