@article{14, keywords = {Photons, Microscopy, Confocal/methods, Fluorescent Dyes/*diagnostic use/metabolism, Calcium/metabolism, Brain Mapping/*methods}, author = {R. Homma and B. Baker and L. Jin and O Garaschuk and A Konnerth and L. Cohen and D. Zecevic}, title = {Wide-field and two-photon imaging of brain activity with voltage- and calcium-sensitive dyes}, abstract = {This review presents three examples of using voltage- or calcium-sensitive dyes to image the activity of the brain. Our aim is to discuss the advantages and disadvantages of each method with particular reference to its application to the study of the brainstem. Two of the examples use wide-field (one-photon) imaging; the third uses two-photon scanning microscopy. Because the measurements have limited signal-to-noise ratio, the paper also discusses the methodological aspects that are critical for optimizing the signal. The three examples are the following. (i) An intracellularly injected voltage-sensitive dye was used to monitor membrane potential in the dendrites of neurons in in vitro preparations. These experiments were directed at understanding how individual neurons convert complex synaptic inputs into the output spike train. (ii) An extracellular, bath application of a voltage-sensitive dye was used to monitor population signals from different parts of the dorsal brainstem. We describe recordings made during respiratory activity. The population signals indicated four different regions with distinct activity correlated with inspiration. (iii) Calcium-sensitive dyes can be used to label many individual cells in the mammalian brain. This approach, combined with two-photon microscopy, made it possible to follow the spike activity in an in vitro brainstem preparation during fictive respiratory rhythms. The organic voltage- and ion-sensitive dyes used today indiscriminatively stain all of the cell types in the preparation. A major effort is underway to develop fluorescent protein sensors of activity for selectively staining individual cell types.}, year = {2009}, journal = {Philos Trans R Soc Lond B Biol Sci}, volume = {364}, edition = {2009/08/05}, number = {1529}, pages = {2453-2467}, month = {Sep 12}, isbn = {1471-2970 (Electronic)0962-8436 (Linking)}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19651647}, note = {Homma, RyotaBaker, Bradley JJin, LeiGaraschuk, OlgaKonnerth, ArthurCohen, Lawrence BZecevic, DejanDC05259/DC/NIDCD NIH HHS/United StatesNS42739/NS/NINDS NIH HHS/United StatesResearch Support, N.I.H., ExtramuralResearch Support, Non-U.S. Gov'tReviewEnglandPhilosophical transactions of the Royal Society of London. Series B, Biological sciencesPhilos Trans R Soc Lond B Biol Sci. 2009 Sep 12;364(1529):2453-67.}, language = {eng}, }