caa a22 4500 606177183 CHVBK 20210128100751.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00018-014-1673-5 doi (NATIONALLICENCE)springer-10.1007/s00018-014-1673-5 Imaging oxygen in neural cell and tissue models by means of anionic cell-permeable phosphorescent nanoparticles [Elektronische Daten] [Ruslan Dmitriev, Sergey Borisov, Alina Kondrashina, Janelle Pakan, Ujval Anilkumar, Jochen Prehn, Alexander Zhdanov, Kieran McDermott, Ingo Klimant, Dmitri Papkovsky] Cell-permeable phosphorescent probes enable the study of cell and tissue oxygenation, bioenergetics, metabolism, and pathological states such as stroke and hypoxia. A number of such probes have been described in recent years, the majority consisting of cationic small molecule and nanoparticle structures. While these probes continue to advance, adequate staining for the study of certain cell types using live imaging techniques remains elusive; this is particularly true for neural cells. Here we introduce novel probes for the analysis of neural cells and tissues: negatively charged poly(methyl methacrylate-co-methacrylic acid)-based nanoparticles impregnated with a phosphorescent Pt(II)-tetrakis(pentafluorophenyl)porphyrin (PtPFPP) dye (this form is referred to as PA1), and with an additional reference/antennae dye poly(9,9-diheptylfluorene-alt-9,9-di-p-tolyl-9H-fluorene) (this form is referred to as PA2). PA1 and PA2 are internalised by endocytosis, result in efficient staining in primary neurons, astrocytes, and PC12 cells and multi-cellular aggregates, and allow for the monitoring of local O2 levels on a time-resolved fluorescence plate reader and PLIM microscope. PA2 also efficiently stains rat brain slices and permits detailed O2 imaging experiments using both one and two-photon intensity-based modes and PLIM modes. Multiplexed analysis of embryonic rat brain slices reveals age-dependent staining patterns for PA2 and a highly heterogeneous distribution of O2 in tissues, which we relate to the localisation of specific progenitor cell populations. Overall, these anionic probes are useful for sensing O2 levels in various cells and tissues, particularly in neural cells, and facilitate high-resolution imaging of O2 in 3D tissue models. Springer Basel, 2014 Biomaterials nationallicence Cell and tissue oxygen nationallicence Intracellular oxygen probe nationallicence Multiplexed detection nationallicence Nanosensors nationallicence Phosphorescence quenching microscopy nationallicence PLIM nationallicence 3D : Three-dimensional nationallicence BLBP : Brain lipid-binding protein nationallicence CPZ : Chlorpromazine nationallicence CTX : Cholera toxin, subunit B nationallicence DIV : Days in vitro nationallicence DMF : N,N-dimethylformamide nationallicence DMSO : Dimethyl sulfoxide nationallicence EIPA : 5-(N-ethyl-N-isopropyl)amiloride nationallicence GFP : Green fluorescent protein nationallicence HBSS : Hanks balanced salt solution nationallicence HXT : Hoechst 33342 nationallicence iO2 : Intracellular O2 nationallicence M β CD : Methyl-β-cyclodextrin nationallicence NP : Nanoparticles nationallicence PA : Polyacrylate NP nationallicence PBS : Phosphate buffered saline nationallicence PDL : Poly-d-lysine nationallicence PDT : Photodynamic therapy nationallicence PLIM : Phosphorescence lifetime imaging microscopy nationallicence PMMA-MA : Poly(methyl methacrylate-co-methacrylic acid nationallicence PFO : Poly(9,9-diheptylfluorene-alt-9,9-di-p-tolyl-9H-fluorene) nationallicence PtPFPP : Pt(II)-tetrakis(pentafluorophenyl)porphine nationallicence ROI : Region of interest nationallicence RT : Room temperature nationallicence TBST : Tris-buffered saline, tween 20 nationallicence TCSPC : Time-correlated single photon counting nationallicence TR-F : Time-resolved fluorescence nationallicence Dmitriev Ruslan School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland aut Borisov Sergey Institute of Analytical Chemistry and Food Chemistry, Graz University of Technology, Stremayrgasse 9, 8010, Graz, Austria aut Kondrashina Alina School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland aut Pakan Janelle Department of Anatomy and Neuroscience, University College Cork, Cork, Ireland aut Anilkumar Ujval Department of Physiology and Medical Physics, Centre for the Study of Neurological Disorders, Royal College of Surgeons in Ireland, Dublin, Ireland aut Prehn Jochen Department of Physiology and Medical Physics, Centre for the Study of Neurological Disorders, Royal College of Surgeons in Ireland, Dublin, Ireland aut Zhdanov Alexander School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland aut McDermott Kieran Department of Anatomy and Neuroscience, University College Cork, Cork, Ireland aut Klimant Ingo Institute of Analytical Chemistry and Food Chemistry, Graz University of Technology, Stremayrgasse 9, 8010, Graz, Austria aut Papkovsky Dmitri School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland aut Cellular and Molecular Life Sciences Springer Basel 72/2(2015-01-01), 367-381 1420-682X 72:2<367 2015 72 18 https://doi.org/10.1007/s00018-014-1673-5 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00018-014-1673-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Dmitriev Ruslan School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland aut NATIONALLICENCE 700 1- Borisov Sergey Institute of Analytical Chemistry and Food Chemistry, Graz University of Technology, Stremayrgasse 9, 8010, Graz, Austria aut NATIONALLICENCE 700 1- Kondrashina Alina School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland aut NATIONALLICENCE 700 1- Pakan Janelle Department of Anatomy and Neuroscience, University College Cork, Cork, Ireland aut NATIONALLICENCE 700 1- Anilkumar Ujval Department of Physiology and Medical Physics, Centre for the Study of Neurological Disorders, Royal College of Surgeons in Ireland, Dublin, Ireland aut NATIONALLICENCE 700 1- Prehn Jochen Department of Physiology and Medical Physics, Centre for the Study of Neurological Disorders, Royal College of Surgeons in Ireland, Dublin, Ireland aut NATIONALLICENCE 700 1- Zhdanov Alexander School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland aut NATIONALLICENCE 700 1- McDermott Kieran Department of Anatomy and Neuroscience, University College Cork, Cork, Ireland aut NATIONALLICENCE 700 1- Klimant Ingo Institute of Analytical Chemistry and Food Chemistry, Graz University of Technology, Stremayrgasse 9, 8010, Graz, Austria aut NATIONALLICENCE 700 1- Papkovsky Dmitri School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland aut NATIONALLICENCE 773 0- Cellular and Molecular Life Sciences Springer Basel 72/2(2015-01-01), 367-381 1420-682X 72:2<367 2015 72 18