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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.” MULTI-PARAMETRIC LIVE CELL MICROSCOPY OF 3D TISSUE MODELS, vol. 1035, Springer International Publishing Ag, 2017, pp. 85–103, doi:10.1007/978-3-319-67358-5_6.
APA
Okkelman, I. A., Foley, T., Papkovsky, D. B., & Dmitriev, R. I. (2017). Multi-Parametric Imaging of Hypoxia and Cell Cycle in Intestinal Organoid Culture. MULTI-PARAMETRIC LIVE CELL MICROSCOPY OF 3D TISSUE MODELS, 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 I. Dmitriev. 2017. “Multi-Parametric Imaging of Hypoxia and Cell Cycle in Intestinal Organoid Culture.” MULTI-PARAMETRIC LIVE CELL MICROSCOPY OF 3D TISSUE MODELS 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 I. Dmitriev. 2017. “Multi-Parametric Imaging of Hypoxia and Cell Cycle in Intestinal Organoid Culture.” MULTI-PARAMETRIC LIVE CELL MICROSCOPY OF 3D TISSUE MODELS 1035: 85–103. doi:10.1007/978-3-319-67358-5_6.
Vancouver
1.
Okkelman IA, Foley T, Papkovsky DB, Dmitriev RI. Multi-Parametric Imaging of Hypoxia and Cell Cycle in Intestinal Organoid Culture. MULTI-PARAMETRIC LIVE CELL MICROSCOPY OF 3D TISSUE MODELS. 2017;1035:85–103.
IEEE
[1]
I. A. Okkelman, T. Foley, D. B. Papkovsky, and R. I. Dmitriev, “Multi-Parametric Imaging of Hypoxia and Cell Cycle in Intestinal Organoid Culture,” MULTI-PARAMETRIC LIVE CELL MICROSCOPY OF 3D TISSUE MODELS, 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 I.},
  isbn         = {978-3-319-67358-5},
  issn         = {0065-2598},
  journal      = {MULTI-PARAMETRIC LIVE CELL MICROSCOPY OF 3D TISSUE MODELS},
  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},
  publisher    = {Springer International Publishing Ag},
  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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