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Multi-Parametric Imaging of Hypoxia and Cell Cycle in Intestinal Organoid Culture

Irina A. Okkelman, Tara Foley, Dmitri B. Papkovsky and Ruslan I. Dmitriev
(2017) MULTI-PARAMETRIC LIVE CELL MICROSCOPY OF 3D TISSUE MODELS. 1035. p.85-103
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

Citation

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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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