| Title: |
ANG-2 for quantitative Na+ determination in living cells by time-resolved fluorescence microscopy |
| Authors: |
Roder, Phillip (M.Sc.); Hille, Carsten (Dr.) |
| Publication Year: |
2014 |
| Collection: |
University of Potsdam: publish.UP |
| Subject Terms: |
ddc:540; Institut für Chemie |
| Description: |
Sodium ions (Na+) play an important role in a plethora of cellular processes, which are complex and partly still unexplored. For the investigation of these processes and quantification of intracellular Na+ concentrations ([Na+]i), two-photon coupled fluorescence lifetime imaging microscopy (2P-FLIM) was performed in the salivary glands of the cockroach Periplaneta americana. For this, the novel Na+-sensitive fluorescent dye Asante NaTRIUM Green-2 (ANG-2) was evaluated, both in vitro and in situ. In this context, absorption coefficients, fluorescence quantum yields and 2P action cross-sections were determined for the first time. ANG-2 was 2P-excitable over a broad spectral range and displayed fluorescence in the visible spectral range. Although the fluorescence decay behaviour of ANG-2 was triexponential in vitro, its analysis indicates a Na+-sensitivity appropriate for recordings in living cells. The Na+-sensitivity was reduced in situ, but the biexponential fluorescence decay behaviour could be successfully analysed in terms of quantitative [Na+]i recordings. Thus, physiological 2P-FLIM measurements revealed a dopamine-induced [Na+]i rise in cockroach salivary gland cells, which was dependent on a Na+-K+-2Cl− cotransporter (NKCC) activity. It was concluded that ANG-2 is a promising new sodium indicator applicable for diverse biological systems. |
| Document Type: |
article in journal/newspaper |
| File Description: |
application/pdf |
| Language: |
English |
| Availability: |
https://publishup.uni-potsdam.de/frontdoor/index/index/docId/7685; https://nbn-resolving.org/urn:nbn:de:kobv:517-opus4-76851; https://publishup.uni-potsdam.de/files/7685/pmnr184.pdf |
| Rights: |
https://creativecommons.org/licenses/by/3.0/ ; info:eu-repo/semantics/openAccess |
| Accession Number: |
edsbas.769D2DE1 |
| Database: |
BASE |