title: Ratiometric Sensing and Imaging of Intracellular pH Using Polyethylenimine-Coated Photon Upconversion Nanoprobes.
abstract: Measurement of changes of pH at various intracellular compartments has potential to solve questions concerning the processing of endocytosed material, regulation of the acidification process, and also acidification of vesicles destined for exocytosis. To monitor these events, the nanosized optical pH probes need to provide ratiometric signals in the optically transparent biological window, target to all relevant intracellular compartments, and to facilitate imaging at subcellular resolution without interference from the biological matrix. To meet these criteria we sensitize the surface conjugated pH sensitive indicator via an upconversion process utilizing an energy transfer from the nanoparticle to the indicator. Live cells were imaged with a scanning confocal microscope equipped with a low-energy 980 nm laser excitation, which facilitated high resolution and penetration depth into the specimen, and low phototoxicity needed for long-term imaging. Our upconversion nanoparticle resonance energy transfer based sensor with polyethylenimine-coating provides high colloidal stability, enhanced cellular uptake, and distribution across cellular compartments. This distribution was modulated with membrane integrity perturbing treatment that resulted into total loss of lysosomal compartments and a dramatic pH shift of endosomal compartments. These nanoprobes are well suited for detection of pH changes in in vitro models with high biological background fluorescence and in in vivo applications, e.g., for the bioimaging of small animal models.
authors.affiliation: Laboratory of Biophysics, Institute of Biomedicine and Medical Research Laboratories, University of Turku , Tykistökatu 6A, 20520 Turku, Finland.|Department of Neurosciences, Karolinska Institutet , von Eulers väg 3, 17177 Stockholm, Sweden.
authors.forename: Tuomas
authors.identifier: 0000-0002-2174-4434
authors.initials: T
authors.lastname: Näreoja
authors.affiliation: Laboratory of Biophysics, Institute of Biomedicine and Medical Research Laboratories, University of Turku , Tykistökatu 6A, 20520 Turku, Finland.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Deguchi
authors.affiliation: Department of Biochemistry/Biotechnology, University of Turku , Tykistökatu 6A, FI-20520 Turku, Finland.
authors.forename: Simon
authors.initials: S
authors.lastname: Christ
authors.affiliation: Department of Biochemistry/Biotechnology, University of Turku , Tykistökatu 6A, FI-20520 Turku, Finland.
authors.forename: Riikka
authors.initials: R
authors.lastname: Peltomaa
authors.affiliation: Pharmaceutical Sciences Laboratory, Faculty of Science and Engineering, Åbo Akademi University , Tykistökatu 6A, 20520 Turku, Finland.
authors.forename: Neeraj
authors.initials: N
authors.lastname: Prabhakar
authors.affiliation: Laboratory of Biophysics, Institute of Biomedicine and Medical Research Laboratories, University of Turku , Tykistökatu 6A, 20520 Turku, Finland.
authors.forename: Elnaz
authors.initials: E
authors.lastname: Fazeli
authors.affiliation: Department of Biochemistry/Biotechnology, University of Turku , Tykistökatu 6A, FI-20520 Turku, Finland.
authors.forename: Niina
authors.initials: N
authors.lastname: Perälä
authors.affiliation: Pharmaceutical Sciences Laboratory, Faculty of Science and Engineering, Åbo Akademi University , Tykistökatu 6A, 20520 Turku, Finland.
authors.forename: Jessica M
authors.identifier: 0000-0001-6085-1112
authors.initials: JM
authors.lastname: Rosenholm
authors.affiliation: Department of Biochemistry/Biotechnology, University of Turku , Tykistökatu 6A, FI-20520 Turku, Finland.
authors.forename: Riikka
authors.identifier: 0000-0002-9930-1676
authors.initials: R
authors.lastname: Arppe
authors.affiliation: Department of Biochemistry/Biotechnology, University of Turku , Tykistökatu 6A, FI-20520 Turku, Finland.
authors.forename: Tero
authors.initials: T
authors.lastname: Soukka
authors.affiliation: BAM - Federal Institute of Materials Research and Testing , Division 1.10 Biophotonics, Richard-Willstätter-Straße 11, 12489 Berlin, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Schäferling
mesh_terms: D045744:Cell Line, Tumor; D005459:Fluorides / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018613:Microscopy, Confocal; D053758:Nanoparticles / Q000737:chemistry* / Q000378:metabolism; D017785:Photons; D011094:Polyethyleneimine / Q000737:chemistry*; D013053:Spectrophotometry; D015019:Yttrium / Q000737:chemistry
chemical_list: C525362:sodium yttriumtetrafluoride; D015019:Yttrium; D011094:Polyethyleneimine; D005459:Fluorides