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Subcellular imaging in the live mouse

Nature Protocols volume 1, pages 775–782 (2006)Cite this article

Abstract

Fluorescent proteins are available in multiple colors and have properties such as intrinsic brightness and high quantum yield that make them optimally suited for in vivo imaging with subcellular resolution in the live mouse. In this protocol, cancer cells in live mice are labeled with green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in the cytoplasm. GFP nuclear labeling is effected by linkage of GFP to histone H2B, and a retroviral vector is used for cytoplasmic labeling with RFP. Double-labeled cells are injected by various methods. High-resolution imaging systems with microscopic optics, in combination with reversible skin flaps over various organs, enable the imaging of dual-color labeled cells at the subcellular level in live animals. The double transfection and selection procedures described here take 6–8 weeks. Cancer cell trafficking, deformation, extravasation, mitosis and cell death can be imaged with clarity.

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Figure 1: Real-time whole-body image of mitotic cell in ear of a live mouse.
Figure 2: Classification of the deformation of HT-1080-dual-color cells in the vessels in the skin.
Figure 3: Migration velocity of HT-1080-dual-color cell in capillaries.
Figure 4: Fate of HCT-116-GFP-RFP human colon cancer cells after portal vein injection.
Figure 5: Fate of MMT GFP-RFP mouse mammary tumor cells after portal vein injection (see Fig. 4 for imaging details).
Figure 6: Intravascular trafficking of HT-1080-GFP-RFP human fibrosarcoma cells.
Figure 7: Time-lapse imaging of extravasation of MMT mouse mammary GFP-RFP tumor cells.
Figure 8: Imaging of extravasation, invasion and proliferation of Lewis lung carcinoma (LLC) GFP-RFP cells.

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Author information

Authors and Affiliations

  1. AntiCancer Inc., 7917 Ostrow Street, San Diego, 92111, California, USA

    Robert M Hoffman & Meng Yang

  2. Dept. of Surgery, University of California, San Diego, 200 West Arbor Drive, San Diego, 92103-8220, CA

    Robert M Hoffman

Authors
  1. Robert M Hoffman
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  2. Meng Yang
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Correspondence to Robert M Hoffman.

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Competing interests

R.M.H. is President of AntiCancer Inc. and M.Y. is a Senior Staff Scientist at AntiCancer Inc. AntiCancer Inc. has commercial activities in the area of fluorescent-protein-based imaging and has a Technology Development Agreement with the Olympus Corporation.

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Hoffman, R., Yang, M. Subcellular imaging in the live mouse. Nat Protoc 1, 775–782 (2006). https://doi.org/10.1038/nprot.2006.109

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