Micropilot: automation of fluorescence microscopy–based imaging for systems biology (original) (raw)

Nature Methods volume 8, pages 246–249 (2011)Cite this article

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Abstract

Quantitative microscopy relies on imaging of large cell numbers but is often hampered by time-consuming manual selection of specific cells. The 'Micropilot' software automatically detects cells of interest and launches complex imaging experiments including three-dimensional multicolor time-lapse or fluorescence recovery after photobleaching in live cells. In three independent experimental setups this allowed us to statistically analyze biological processes in detail and is thus a powerful tool for systems biology.

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Acknowledgements

This study was technically supported by the use of the European Molecular Biology Laboratory Advanced Light Microscopy Facility and Information Technology Service Unit. We acknowledge C. Chapuis for the CBX1-EGFP BAC cell line expressing H2B-mCherry. This project was funded by grants to J.E. and R.P. (within the MitoCheck, MitoSys and Systems Microscopy consortia) by the European Commission (LSHG-CT-2004-503464, FP7/2007-2013-241548 and FP7/2007-2013-258068), to R.P. by the Landesstiftung Baden-Württemberg in the framework of the research program RNS/RNAi and within the Nationales Genomforschungsnetz-Plus consortium IG-CSG (01GS0865). T.H.T. is financed by the Deutsche Forschungsgemeinschaft Graduiertenkolleg GRK118.

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Author notes

  1. Urban Liebel
    Present address: Present address: Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen, Germany.,

Authors and Affiliations

  1. Advanced Light Microscopy Facility, European Molecular Biology Laboratory, Heidelberg, Germany
    Christian Conrad, Jutta Bulkescher & Rainer Pepperkok
  2. Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    Annelie Wünsche, Tze Heng Tan, Fatima Verissimo, Thomas Walter, Urban Liebel, Rainer Pepperkok & Jan Ellenberg
  3. Leica Microsystems GmbH, Mannheim, Germany
    Frank Sieckmann
  4. Department of Cellular and Molecular Pharmacology, University of California, San Francisco, California, USA
    Arthur Edelstein

Authors

  1. Christian Conrad
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  2. Annelie Wünsche
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  3. Tze Heng Tan
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  4. Jutta Bulkescher
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  5. Frank Sieckmann
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  6. Fatima Verissimo
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  7. Arthur Edelstein
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  8. Thomas Walter
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  9. Urban Liebel
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  10. Rainer Pepperkok
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  11. Jan Ellenberg
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Contributions

C.C. developed the 'Micropilot' software and drafted the manuscript. A.W. developed the Visual Basic for Applications macro and performed and analyzed the automatic FRAP experiments. T.H.T. developed the feature selection, extended classification to multiple channels and acquired and analyzed the ERES images. F.V. performed the ERES experiments. J.B. performed and analyzed the spindle length experiments. F.S. and U.L. developed the computer-aided microscopy interface and set up software prototypes. A.E. developed the communication of μManager with Micropilot. T.W. helped with image processing and object feature design. R.P. supervised the project. J.E. supervised the project and revised the manuscript.

Corresponding authors

Correspondence toRainer Pepperkok or Jan Ellenberg.

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Competing interests

F.S and U.L. filed a patent application covering the CAM approach (Patent Cooperation Treaty/European Patent 2007/059351/US patent application 20100103253). F.S. is employed by Leica Microsystems.

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Conrad, C., Wünsche, A., Tan, T. et al. Micropilot: automation of fluorescence microscopy–based imaging for systems biology.Nat Methods 8, 246–249 (2011). https://doi.org/10.1038/nmeth.1558

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