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Bioorthogonal Probes for Polo-like Kinase 1 Imaging and Quantification

Authors

  • Dr. Ghyslain Budin,

    1. Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114 (USA)
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    • These authors contributed equally to this work.

  • Dr. Katherine S. Yang,

    1. Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114 (USA)
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    • These authors contributed equally to this work.

  • Dr. Thomas Reiner,

    1. Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114 (USA)
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  • Prof. Ralph Weissleder

    Corresponding author
    1. Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114 (USA)
    2. Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115 (USA)
    • Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114 (USA)
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  • This work was supported by the National Institutes of Health (NIH) grant number RO1EB010011, K.S. was supported by a NIH grant T32-CA79443 and T.R. was supported by a grant from the German Academy of Sciences Leopoldina (LPDS 2009-24). We thank Joshua Dunham and Alex Zaltsman for image processing, and Dr. Robert Yang and Prof. Peter Sorger for assistance with cellWoRx and Image Rail.

Abstract

original image

Click inside: A nuclear protein target, polo-like kinase 1 (PLK1) was imaged using a biocompatible bioorthogonal ligation between a specific drug and a fluorescent dye in live cells (see picture). Colocalization of the dye and the protein target was confirmed by antibody staining and by expressing a GFP construct of PLK1. The two-step PLK1 imaging procedure was used to quantify PLK1 expression levels in cancer cell lines of various tissue origins.

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