PMID- 19255672
OWN - NLM
STAT- MEDLINE
DCOM- 20090520
LR  - 20090303
IS  - 1474-905X (Print)
IS  - 1474-905X (Linking)
VI  - 8
IP  - 3
DP  - 2009 Mar
TI  - Two-photon fluorescence lifetime imaging of intracellular chloride in cockroach 
      salivary glands.
PG  - 319-27
LID - 10.1039/b813797h [doi]
AB  - Although chloride plays an important role in many cellular processes, there is a 
      lack of data about intracellular chloride concentrations [Cl(-)](i), particularly 
      due to technical problems. To overcome that, in this study fluorescence lifetime 
      imaging microscopy in the time-domain by using time-correlated single-photon 
      counting was combined with two-photon excitation (2P-FLIM). This 2P-FLIM setup 
      has been successfully used with the Cl(-)-sensitive fluorescent dye 
      N-(ethoxycarbonylmethyl)-6-methoxy-quinolinium bromide (MQAE) in order to measure 
      [Cl(-)](i) in cockroach salivary glands, a well-established model system for 
      studying epithelial ion transport processes. MQAE was well suitable for 
      two-photon excitation, when loaded into cells, and displayed a sufficient dynamic 
      range of its fluorescence decay time changes in response to variation of 
      [Cl(-)](i) according to the Stern-Volmer relationship. On this basis a uniform 
      [Cl(-)](i) in the range of 42-80 mM with a mean value of 59 mM +/- 1 mM was found 
      in resting cockroach salivary ducts, indicating active Cl(-) accumulation. 
      However, exposure to Cl(-)-free saline caused only a moderate [Cl(-)](i) drop to 
      48 mM +/- 4 mM, suggesting a relatively low basolateral Cl(-) permeability in 
      ducts, at least under resting conditions. Additionally, bath application of the 
      biogenic amine dopamine, known to stimulate the saliva modification in the ducts, 
      caused no significant [Cl(-)](i) changes. These results suggest a more complex 
      scenario of [Cl(-)](i) homeostasis in cockroach salivary ducts. In conclusion, 
      2P-FLIM seems to be a suitable technique for quantitative [Cl(-)](i) measurements 
      in many biological systems.
FAU - Hille, Carsten
AU  - Hille C
AD  - Physical Chemistry/Applied Laser Sensing, Institute of Chemistry & 
      Interdisciplinary Center for Photonics, University of Potsdam, 
      Karl-Liebknecht-Str. 24-25, 14476, Potsdam-Golm, Germany. hille@uni-potsdam.de
FAU - Lahn, Mattes
AU  - Lahn M
FAU - Lohmannsroben, Hans-Gerd
AU  - Lohmannsroben HG
FAU - Dosche, Carsten
AU  - Dosche C
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090105
PL  - England
TA  - Photochem Photobiol Sci
JT  - Photochemical & photobiological sciences : Official journal of the European 
      Photochemistry Association and the European Society for Photobiology
JID - 101124451
RN  - 0 (Chlorides)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Quinolinium Compounds)
RN  - 18010-05-4 (N-(ethoxycarbonylmethyl)-6-methoxyquinolinium)
SB  - IM
MH  - Animals
MH  - Chlorides/*analysis
MH  - Cockroaches/chemistry/radiation effects
MH  - Fluorescent Dyes/chemistry
MH  - Microscopy, Fluorescence, Multiphoton/*methods
MH  - Photons
MH  - Quinolinium Compounds/chemistry
MH  - Salivary Glands/*chemistry/radiation effects
EDAT- 2009/03/04 09:00
MHDA- 2009/05/21 09:00
CRDT- 2009/03/04 09:00
PHST- 2009/03/04 09:00 [entrez]
PHST- 2009/03/04 09:00 [pubmed]
PHST- 2009/05/21 09:00 [medline]
AID - 10.1039/b813797h [doi]
PST - ppublish
SO  - Photochem Photobiol Sci. 2009 Mar;8(3):319-27. doi: 10.1039/b813797h. Epub 2009 
      Jan 5.