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Fluorescence Microscopy for ATP Internalization Mediated by Macropinocytosis in Human Tumor Cells and Tumor-xenografted Mice
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1Department of Biological Sciences,Ohio University, 2Molecular and Cellular Biology Program,Ohio University, 3Neuroscience Program,Ohio University, 4Edison Biotechnology Institute,Ohio University, 5Translational Biomedical Sciences Program,Ohio University, 6Honors Tutorial College,Ohio University, 7Department of Biomedical Sciences, Heritage College of Osteopathic Medicine,Ohio University
Chapters
- 00:05Introduction
- 01:05Tumor Cell Culture and Xenograft Tumor Development
- 02:41HMWFD and NHF-ATP Solution Preparation
- 03:21ATP Internalization in Tumor Xenograft
- 03:53Cryo-embedding
- 04:45Tissue Slide Fixation
- 05:25Fluorescence Microscopy and Image Acquisition
- 07:12Results: Assessment of Nonhydrolyzable Fluorescent ATP Internalization
- 07:44Conclusion
We developed a reproducible method to visualize the internalization of nonhydrolyzable fluorescent adenosine triphosphate (ATP), an ATP surrogate, with high cellular resolution. We validated our method using independent in vitro and in vivo assays-human tumor cell lines and immunodeficient mice xenografted with human tumor tissue.
