Time-domain fluorescence lifetime imaging for intracellular pH sensing in living tissues.
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.
Hille
Carsten
C
Institute of Chemistry, University of Potsdam, Karl-Liebknecht-Str. 24-25, 14476, Potsdam-Golm, Germany.
Berg
Maik
M
Bressel
Lena
L
Munzke
Dorit
D
Primus
Philipp
P
Löhmannsröben
Hans-Gerd
HG
Dosche
Carsten
C
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
D005452:Fluoresceins; D005456:Fluorescent Dyes; C043829:2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein