Imaging of oxygenation in 3D tissue models with multi-modal phosphorescent probes

Cell-penetrating phosphorescence based probes allow real-time, high-resolution imaging of O2 concentration in respiring cells and 3D tissue models. We have developed a panel of such probes, small molecule and nanoparticle structures, which have different spectral characteristics, cell penetrating and tissue staining behavior. The probes are compatible with conventional live cell imaging platforms and can be used in different detection modalities, including ratiometric intensity and PLIM (Phosphorescence Lifetime IMaging) under one- or two-photon excitation. Analytical performance of these probes and utility of the O2 imaging method have been demonstrated with different types of samples: 2D cell cultures, multi-cellular spheroids from cancer cell lines and primary neurons, excised slices from mouse brain, colon and bladder tissue, and live animals. They are particularly useful for hypoxia research, ex-vivo studies of tissue physiology, cell metabolism, cancer, inflammation, and multiplexing with many conventional fluorophors and markers of cellular function.

[1]  Alexander V. Zhdanov,et al.  Imaging of neurosphere oxygenation with phosphorescent probes. , 2013, Biomaterials.

[2]  Ingo Klimant,et al.  Intracellular O2 sensing probe based on cell-penetrating phosphorescent nanoparticles. , 2011, ACS nano.

[3]  Christopher S. Burke,et al.  Peptide-bridged dinuclear Ru(II) complex for mitochondrial targeted monitoring of dynamic changes to oxygen concentration and ROS generation in live mammalian cells. , 2014, Journal of the American Chemical Society.

[4]  Aamir A. Khan,et al.  Easily prepared ruthenium-complex nanomicelle probes for two-photon quantitative imaging of oxygen in aqueous media , 2015 .

[5]  Alexander V. Zhdanov,et al.  A Phosphorescent Nanoparticle‐Based Probe for Sensing and Imaging of (Intra)Cellular Oxygen in Multiple Detection Modalities , 2012 .

[6]  Dmitri B Papkovsky,et al.  Assessment of cellular oxygen gradients with a panel of phosphorescent oxygen-sensitive probes. , 2012, Analytical chemistry.

[7]  Dmitri B Papkovsky,et al.  Biological detection by optical oxygen sensing. , 2013, Chemical Society reviews.

[8]  S. Takayama,et al.  Opportunities and challenges for use of tumor spheroids as models to test drug delivery and efficacy. , 2012, Journal of controlled release : official journal of the Controlled Release Society.

[9]  Alexander V. Zhdanov,et al.  Small molecule phosphorescent probes for O2 imaging in 3D tissue models. , 2014, Biomaterials science.

[10]  J. Pakan,et al.  Imaging oxygen in neural cell and tissue models by means of anionic cell-permeable phosphorescent nanoparticles , 2014, Cellular and Molecular Life Sciences.

[11]  Alexander V. Zhdanov,et al.  Analysis of intracellular oxygen and metabolic responses of mammalian cells by time-resolved fluorometry. , 2007, Analytical chemistry.

[12]  R. Erzurumlu,et al.  In vivo imaging of brain metabolism activity using a phosphorescent oxygen-sensitive probe , 2013, Journal of Neuroscience Methods.

[13]  Dmitri B Papkovsky,et al.  Intracellular oxygen-sensitive phosphorescent probes based on cell-penetrating peptides. , 2010, Analytical biochemistry.

[14]  R. I. Dmitriev,et al.  Intracellular probes for imaging oxygen concentration: how good are they? , 2015, Methods and applications in fluorescence.

[15]  O. Wolfbeis,et al.  Optical methods for sensing and imaging oxygen: materials, spectroscopies and applications. , 2014, Chemical Society reviews.

[16]  Dmitri B. Papkovsky,et al.  Monitoring of cell oxygenation and responses to metabolic stimulation by intracellular oxygen sensing technique , 2017 .

[17]  Ruslan I. Dmitriev,et al.  Optical probes and techniques for O2 measurement in live cells and tissue , 2012, Cellular and Molecular Life Sciences.

[18]  John W. Haycock,et al.  Long-lived metal complexes open up microsecond lifetime imaging microscopy under multiphoton excitation: from FLIM to PLIM and beyond , 2014 .

[19]  F. Pampaloni,et al.  The third dimension bridges the gap between cell culture and live tissue , 2007, Nature Reviews Molecular Cell Biology.

[20]  E. Betzig,et al.  Noninvasive Imaging of 3 D Dynamics in Thickly Fluorescent Specimens Beyond the Diffraction Limit , 2013 .

[21]  Ingo Klimant,et al.  Versatile Conjugated Polymer Nanoparticles for High-Resolution O2 Imaging in Cells and 3D Tissue Models. , 2015, ACS nano.

[22]  W. Becker Fluorescence lifetime imaging – techniques and applications , 2012, Journal of microscopy.

[23]  I. Slutsky,et al.  Spatially resolved recording of transient fluorescence‐lifetime effects by line‐scanning TCSPC , 2014, Microscopy research and technique.

[24]  A. Dale,et al.  Frontiers in Optical Imaging of Cerebral Blood Flow and Metabolism , 2012, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[25]  O. Wolfbeis,et al.  Targetable phosphorescent oxygen nanosensors for the assessment of tumor mitochondrial dysfunction by monitoring the respiratory activity. , 2014, Angewandte Chemie.

[26]  R. I. Dmitriev,et al.  Multi-parametric O₂ imaging in three-dimensional neural cell models with the phosphorescent probes. , 2015, Methods in molecular biology.

[27]  M. Davidson,et al.  Noninvasive Imaging beyond the Diffraction Limit of 3D Dynamics in Thickly Fluorescent Specimens , 2012, Cell.