title: Time-domain fluorescence lifetime imaging for intracellular pH sensing in living tissues.
abstract: pH sensing in living cells represents one of the most prominent topics in biochemistry and physiology. In this study we performed one-photon and two-photon time-domain fluorescence lifetime imaging with a laser-scanning microscope using the time-correlated single-photon counting technique for imaging intracellular pH levels. The suitability of different commercial fluorescence dyes for lifetime-based pH sensing is discussed on the basis of in vitro as well of in situ measurements. Although the tested dyes are suitable for intensity-based ratiometric measurements, for lifetime-based techniques in the time-domain so far only BCECF seems to meet the requirements of reliable intracellular pH recordings in living cells.
authors.affiliation: Institute of Chemistry, University of Potsdam, Karl-Liebknecht-Str. 24-25, 14476, Potsdam-Golm, Germany.
authors.forename: Carsten
authors.initials: C
authors.lastname: Hille
authors.forename: Maik
authors.initials: M
authors.lastname: Berg
authors.forename: Lena
authors.initials: L
authors.lastname: Bressel
authors.forename: Dorit
authors.initials: D
authors.lastname: Munzke
authors.forename: Philipp
authors.initials: P
authors.lastname: Primus
authors.forename: Hans-Gerd
authors.initials: HG
authors.lastname: Löhmannsröben
authors.forename: Carsten
authors.initials: C
authors.lastname: Dosche
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D003952:Diagnostic Imaging / Q000379:methods*; D004727:Endothelium / Q000166:cytology*; D005452:Fluoresceins / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007834:Lasers; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods*; D017785:Photons; D013997:Time Factors
chemical_list: D005452:Fluoresceins; D005456:Fluorescent Dyes; C043829:2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein