Imaging oxygen in neural cell and tissue models by means of anionic cell-permeable phosphorescent nanoparticles

Ruslan I. Dmitriev*, Sergey M. Borisov, Alina V. Kondrashina, Janelle M. P. Pakan, Ujval Anilkumar, Jochen H. M. Prehn, Alexander V. Zhdanov, Kieran W. McDermott, Ingo Klimant, Dmitri B. Papkovsky

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

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 O-2 levels on a time-resolved fluorescence plate reader and PLIM microscope. PA2 also efficiently stains rat brain slices and permits detailed O-2 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 O-2 in tissues, which we relate to the localisation of specific progenitor cell populations. Overall, these anionic probes are useful for sensing O-2 levels in various cells and tissues, particularly in neural cells, and facilitate high-resolution imaging of O-2 in 3D tissue models.

Original languageEnglish
Pages (from-to)367-381
Number of pages15
JournalCellular and Molecular Life Sciences
Volume72
Issue number2
DOIs
Publication statusPublished - Jan 2015

Keywords / Materials (for Non-textual outputs)

  • Biomaterials
  • Cell and tissue oxygen
  • Intracellular oxygen probe
  • Multiplexed detection
  • Nanosensors
  • Phosphorescence quenching microscopy
  • PLIM
  • INTRACELLULAR OXYGEN
  • IN-VIVO
  • ORGANOTYPIC CULTURES
  • SLICE CULTURES
  • MOUSE MODELS
  • LIVE CELLS
  • BLOOD-FLOW
  • PROBES
  • SENSORS
  • O-2

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