Journal
CELLULAR AND MOLECULAR LIFE SCIENCES
Volume 72, Issue 2, Pages 367-381Publisher
SPRINGER BASEL AG
DOI: 10.1007/s00018-014-1673-5
Keywords
Biomaterials; Cell and tissue oxygen; Intracellular oxygen probe; Multiplexed detection; Nanosensors; Phosphorescence quenching microscopy; PLIM
Categories
Funding
- Science Foundation Ireland [12/RC/2276]
- European Commission [FP7-HEALTH-2012-INNOVATION-304842-2]
- Irish Research Council for Science, Engineering and Technology
- Health Research Board
- Programme for Research at Third Level Institutions Cycle 4
- Marie Curie IAPP Oxy-Sense [230641]
Ask authors/readers for more resources
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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available