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Multi-parametric imaging of hypoxia and cell cycle in intestinal organoid culture

Author
Organization
Abstract
Dynamics of oxygenation of tissue and stem cell niches are important for understanding physiological function of the intestine in normal and diseased states. Only a few techniques allow live visualization of tissue hypoxia at cellular level and in three dimensions. We describe an optimized protocol, which uses cell-penetrating O-2-sensitive probe, Pt-Glc and phosphorescence lifetime imaging microscopy (PLIM), to analyze O-2 distribution in mouse intestinal organoids. Unlike the other indirect and end-point hypoxia stains, or point measurements with microelectrodes, this method provides high-resolution real-time visualization of O-2 in organoids. Multiplexing with conventional fluorescent live cell imaging probes such as the Hoechst 33342-based FLIM assay of cell proliferation, and immunofluorescence staining of endogenous proteins, allows analysis of key physiologic parameters under O-2 control in organoids. The protocol is useful for gastroenterology and physiology of intestinal tissue, hypoxia research, regenerative medicine, studying host-microbiota interactions and bioenergetics.
Keywords
3D TISSUE MODELS, PHOSPHORESCENT PROBES, STEM-CELLS, IN-VITRO, OXYGEN, VIVO, FLUORESCENCE, MICROBIOTA, METAGENOME, MICROSCOPY, Cell cycle, FLIM, Intestinal organoids, Live cell microscopy, Oxygen, Phosphorescence quenching, PLIM

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MLA
Okkelman, Irina A., et al. “Multi-Parametric Imaging of Hypoxia and Cell Cycle in Intestinal Organoid Culture.” Advances in Experimental Medicine and Biology, vol. 1035, 2017, pp. 85–103, doi:10.1007/978-3-319-67358-5_6.
APA
Okkelman, I. A., Foley, T., Papkovsky, D. B., & Dmitriev, R. (2017). Multi-parametric imaging of hypoxia and cell cycle in intestinal organoid culture. Advances in Experimental Medicine and Biology, 1035, 85–103. https://doi.org/10.1007/978-3-319-67358-5_6
Chicago author-date
Okkelman, Irina A., Tara Foley, Dmitri B. Papkovsky, and Ruslan Dmitriev. 2017. “Multi-Parametric Imaging of Hypoxia and Cell Cycle in Intestinal Organoid Culture.” Advances in Experimental Medicine and Biology 1035: 85–103. https://doi.org/10.1007/978-3-319-67358-5_6.
Chicago author-date (all authors)
Okkelman, Irina A., Tara Foley, Dmitri B. Papkovsky, and Ruslan Dmitriev. 2017. “Multi-Parametric Imaging of Hypoxia and Cell Cycle in Intestinal Organoid Culture.” Advances in Experimental Medicine and Biology 1035: 85–103. doi:10.1007/978-3-319-67358-5_6.
Vancouver
1.
Okkelman IA, Foley T, Papkovsky DB, Dmitriev R. Multi-parametric imaging of hypoxia and cell cycle in intestinal organoid culture. Advances in Experimental Medicine and Biology. 2017;1035:85–103.
IEEE
[1]
I. A. Okkelman, T. Foley, D. B. Papkovsky, and R. Dmitriev, “Multi-parametric imaging of hypoxia and cell cycle in intestinal organoid culture,” Advances in Experimental Medicine and Biology, vol. 1035, pp. 85–103, 2017.
@article{8687873,
  abstract     = {{Dynamics of oxygenation of tissue and stem cell niches are important for understanding physiological function of the intestine in normal and diseased states. Only a few techniques allow live visualization of tissue hypoxia at cellular level and in three dimensions. We describe an optimized protocol, which uses cell-penetrating O-2-sensitive probe, Pt-Glc and phosphorescence lifetime imaging microscopy (PLIM), to analyze O-2 distribution in mouse intestinal organoids. Unlike the other indirect and end-point hypoxia stains, or point measurements with microelectrodes, this method provides high-resolution real-time visualization of O-2 in organoids. Multiplexing with conventional fluorescent live cell imaging probes such as the Hoechst 33342-based FLIM assay of cell proliferation, and immunofluorescence staining of endogenous proteins, allows analysis of key physiologic parameters under O-2 control in organoids. The protocol is useful for gastroenterology and physiology of intestinal tissue, hypoxia research, regenerative medicine, studying host-microbiota interactions and bioenergetics.}},
  author       = {{Okkelman, Irina A. and Foley, Tara and Papkovsky, Dmitri B. and Dmitriev, Ruslan}},
  isbn         = {{9783319673578}},
  issn         = {{0065-2598}},
  journal      = {{Advances in Experimental Medicine and Biology}},
  keywords     = {{3D TISSUE MODELS,PHOSPHORESCENT PROBES,STEM-CELLS,IN-VITRO,OXYGEN,VIVO,FLUORESCENCE,MICROBIOTA,METAGENOME,MICROSCOPY,Cell cycle,FLIM,Intestinal organoids,Live cell microscopy,Oxygen,Phosphorescence quenching,PLIM}},
  language     = {{eng}},
  pages        = {{85--103}},
  title        = {{Multi-parametric imaging of hypoxia and cell cycle in intestinal organoid culture}},
  url          = {{http://dx.doi.org/10.1007/978-3-319-67358-5_6}},
  volume       = {{1035}},
  year         = {{2017}},
}

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