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Multi-parametric O2 Imaging in Three-Dimensional Neural Cell Models with the Phosphorescent Probes

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1254)

Abstract

Recent progress in bio-imaging has allowed detailed mechanistic studies of neural cell function in complex 3D tissue models including multicellular aggregates , neurospheres , excised brain slices , ganglia , and organoids . Molecular oxygen (O2 ) is an important metabolite and an environmental parameter which determines the viability and physiological status of neural cells within tissue. Here we describe standard method for monitoring O2 in 3D tissue models using phosphorescence lifetime imaging microscopy (PLIM ) and cell-penetrating O2-sensing probes . The O2 probes can be multiplexed with many conventional fluorescence based live cell biomarkers and also end-point immunofluorescence staining . The multi-parametric O2 imaging method is particularly useful for areas such as stem cell development and differentiation , hypoxia research, neurodegenerative disorders , regeneration of brain tissue, evaluation of new drugs , and development of novel tissue models.

Key words

High-resolution microscopy Imaging Oxygen Phosphorescence quenching PLIM 
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Notes

Acknowledgments

This work was supported by the Science Foundation Ireland, grants 12/RC/2276 and 12/TIDA/B2413, and the EC FP7 Program, grant NanoBio4Trans (No. 304842-2). We thank Dr. J. M.P. Pakan and Dr. Y.M. Nolan (Department of Anatomy and Neuroscience, University College Cork) for the help in preparation of brain slices and neurosphere cultures.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.School of Biochemistry and Cell BiologyUniversity College CorkCorkIreland

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