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

被引：14

作者：

Okkelman, Irina A. ^{[1
]}

Foley, Tara ^{[2
]}

Papkovsky, Dmitri B. ^{[1
]}

Dmitriev, Ruslan I. ^{[3
]}

机构：

[1] Univ Coll Cork, Lab Biophys & Bioanal, Sch Biochem & Cell Biol, Cork, Ireland

[2] Univ Coll Cork, Dept Anat & Neurosci, Cork, Ireland

[3] Univ Coll Cork, Sch Biochem & Cell Biol, Metab Imaging Grp, Cork, Ireland

来源：

MULTI-PARAMETRIC LIVE CELL MICROSCOPY OF 3D TISSUE MODELS | 2017年 / 1035卷

关键词：

Cell cycle; FLIM; Intestinal organoids; Live cell microscopy; Oxygen; Phosphorescence quenching; PLIM; IN-VITRO; OXYGEN; PROBES; FLUORESCENCE; METAGENOME; MICROBIOTA; FIBROSIS; HEALTHY; COLON;

D O I：

10.1007/978-3-319-67358-5_6

中图分类号：

Q813 [细胞工程];

学科分类号：

摘要：

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.

引用

页码：85 / 103

页数：19

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