@article{Marandi2002,
	title = {Two-photon chloride imaging in neurons of brain slices.},
	volume = {445},
	issn = {0031-6768},
	url = {http://www.ncbi.nlm.nih.gov/pubmed/12466938},
	doi = {10.1007/s00424-002-0933-7},
	abstract = {Two-photon laser scanning microscopy has been used successfully for imaging activity-dependent changes of intracellular calcium and sodium levels. Here we introduce a simple technique for two-photon chloride imaging in intact neurons. It involves the use of the membrane-permeable Cl(-) indicator dye MQAE [N-(6-methoxyquinolyl) acetoethyl ester]. Brief incubation with MQAE produced a robust loading of cells in slices from various brain regions including hippocampus, cortex and cerebellum. In contrast to conventional fluorescence measurements using MQAE, two-photon imaging was not affected in a major way by dye bleaching and phototoxic damage. Instead, it allowed prolonged recordings of time-resolved changes in intracellular chloride concentration in somata and dendrites. As an example of an application we imaged GABA-mediated Cl(-) transients in pyramidal cells of cortical and hippocampal slices as well as in cerebellar Purkinje neurons. By combining Cl(-) imaging with the gramicidin-based perforated-patch-clamp technique we showed that changes in MQAE fluorescence are proportional to the magnitudes of GABA-evoked transmembrane Cl(-) fluxes. Thus, MQAE-based two-photon microscopy promises to be a valuable technique for many applications requiring chloride imaging in single cells.},
	number = {3},
	urldate = {2012-07-30},
	journal = {Pflügers Archiv : European journal of physiology},
	author = {Marandi, Nima and Konnerth, Arthur and Garaschuk, Olga},
	month = dec,
	year = {2002},
	pmid = {12466938},
	keywords = {Animals, Brain, Brain: metabolism, Chemical, Chlorides, Chlorides: metabolism, Confocal, Confocal: methods, Fluorescent Dyes, Intracellular Membranes, Intracellular Membranes: metabolism, Microscopy, Models, Neurons, Neurons: metabolism, Photons, Quinolines, Rats, Sprague-Dawley},
	pages = {357--65},
}

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In contrast to conventional fluorescence measurements using MQAE, two-photon imaging was not affected in a major way by dye bleaching and phototoxic damage. Instead, it allowed prolonged recordings of time-resolved changes in intracellular chloride concentration in somata and dendrites. As an example of an application we imaged GABA-mediated Cl(-) transients in pyramidal cells of cortical and hippocampal slices as well as in cerebellar Purkinje neurons. By combining Cl(-) imaging with the gramicidin-based perforated-patch-clamp technique we showed that changes in MQAE fluorescence are proportional to the magnitudes of GABA-evoked transmembrane Cl(-) fluxes. Thus, MQAE-based two-photon microscopy promises to be a valuable technique for many applications requiring chloride imaging in single cells.","number":"3","urldate":"2012-07-30","journal":"Pflügers Archiv : European journal of physiology","author":[{"propositions":[],"lastnames":["Marandi"],"firstnames":["Nima"],"suffixes":[]},{"propositions":[],"lastnames":["Konnerth"],"firstnames":["Arthur"],"suffixes":[]},{"propositions":[],"lastnames":["Garaschuk"],"firstnames":["Olga"],"suffixes":[]}],"month":"December","year":"2002","pmid":"12466938","keywords":"Animals, Brain, Brain: metabolism, Chemical, Chlorides, Chlorides: metabolism, Confocal, Confocal: methods, Fluorescent Dyes, Intracellular Membranes, Intracellular Membranes: metabolism, Microscopy, Models, Neurons, Neurons: metabolism, Photons, Quinolines, Rats, Sprague-Dawley","pages":"357–65","bibtex":"@article{Marandi2002,\n\ttitle = {Two-photon chloride imaging in neurons of brain slices.},\n\tvolume = {445},\n\tissn = {0031-6768},\n\turl = {http://www.ncbi.nlm.nih.gov/pubmed/12466938},\n\tdoi = {10.1007/s00424-002-0933-7},\n\tabstract = {Two-photon laser scanning microscopy has been used successfully for imaging activity-dependent changes of intracellular calcium and sodium levels. Here we introduce a simple technique for two-photon chloride imaging in intact neurons. It involves the use of the membrane-permeable Cl(-) indicator dye MQAE [N-(6-methoxyquinolyl) acetoethyl ester]. Brief incubation with MQAE produced a robust loading of cells in slices from various brain regions including hippocampus, cortex and cerebellum. In contrast to conventional fluorescence measurements using MQAE, two-photon imaging was not affected in a major way by dye bleaching and phototoxic damage. Instead, it allowed prolonged recordings of time-resolved changes in intracellular chloride concentration in somata and dendrites. As an example of an application we imaged GABA-mediated Cl(-) transients in pyramidal cells of cortical and hippocampal slices as well as in cerebellar Purkinje neurons. By combining Cl(-) imaging with the gramicidin-based perforated-patch-clamp technique we showed that changes in MQAE fluorescence are proportional to the magnitudes of GABA-evoked transmembrane Cl(-) fluxes. 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