{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T05:48:04Z","timestamp":1791006484834,"version":"4.1.0"},"reference-count":39,"publisher":"Society for Neuroscience","issue":"4","license":[{"start":{"date-parts":[[2004,7,28]],"date-time":"2004-07-28T00:00:00Z","timestamp":1090972800000},"content-version":"vor","delay-in-days":182,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-sa\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J. Neurosci."],"published-print":{"date-parts":[[2004,1,28]]},"abstract":"<jats:p>Nonlinear microscopy has proven to be essential for neuroscience investigations of thick tissue preparations. However, the optical recording of fast (\u223c1 msec) cellular electrical activity has never until now been successfully combined with this imaging modality. Through the use of second-harmonic generation microscopy of primary<jats:italic>Aplysia<\/jats:italic>neurons in culture labeled with 4-[4-(dihexylamino)phenyl][ethynyl]-1-(4-sulfobutyl)pyridinium (inner salt), we optically recorded action potentials with 0.833 msec temporal and 0.6 \u03bcm spatial resolution on soma and neurite membranes. Second-harmonic generation response as a function of change in membrane potential was found to be linear with a signal change of \u223c6%\/100 mV. The signal-to-noise ratio was \u223c1 for single-trace action potential recordings but was readily increased to \u223c6\u20137 with temporal averaging of \u223c50 scans. Photodamage was determined to be negligible by observing action potential characteristics, cellular resting potential, and gross cellular morphology during and after laser illumination. High-resolution (micrometer scale) optical recording of membrane potential activity by previous techniques has been limited to imaging depths an order of magnitude less than nonlinear methods. Because second-harmonic generation is capable of imaging up to \u223c400 \u03bcm deep into intact tissue with submicron resolution and little out-of-focus photodamage or bleaching, its ability to record fast electrical activity should prove valuable to future electrophysiology studies.<\/jats:p>","DOI":"10.1523\/jneurosci.4840-03.2004","type":"journal-article","created":{"date-parts":[[2004,1,29]],"date-time":"2004-01-29T03:23:41Z","timestamp":1075346621000},"page":"999-1003","source":"Crossref","is-referenced-by-count":146,"title":["Optical Recording of Action Potentials with Second-Harmonic Generation Microscopy"],"prefix":"10.1523","volume":"24","author":[{"given":"Daniel A.","family":"Dombeck","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mireille","family":"Blanchard-Desce","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Watt W.","family":"Webb","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"393","published-online":{"date-parts":[[2004,1,28]]},"reference":[{"key":"2023041303045781000_24.4.999.1","doi-asserted-by":"crossref","first-page":"1615","DOI":"10.1152\/jn.1999.82.3.1615","volume":"82","year":"1999","journal-title":"J Neurophysiol"},{"key":"2023041303045781000_24.4.999.2","doi-asserted-by":"crossref","first-page":"1371","DOI":"10.1152\/jn.1998.79.3.1371","volume":"79","year":"1998","journal-title":"J Neurophysiol"},{"key":"2023041303045781000_24.4.999.3","doi-asserted-by":"crossref","first-page":"1616","DOI":"10.1016\/S0006-3495(96)79365-7","volume":"71","year":"1996","journal-title":"Biophys J"},{"key":"2023041303045781000_24.4.999.4","doi-asserted-by":"crossref","first-page":"672","DOI":"10.1016\/S0006-3495(93)81126-3","volume":"65","year":"1993","journal-title":"Biophys J"},{"key":"2023041303045781000_24.4.999.5","doi-asserted-by":"crossref","first-page":"2272","DOI":"10.1016\/S0006-3495(99)77383-2","volume":"76","year":"1999","journal-title":"Biophys J"},{"key":"2023041303045781000_24.4.999.6","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1016\/S0006-3495(97)78086-X","volume":"73","year":"1997","journal-title":"Biophys J"},{"key":"2023041303045781000_24.4.999.7","doi-asserted-by":"crossref","first-page":"3341","DOI":"10.1016\/S0006-3495(99)77165-1","volume":"77","year":"1999","journal-title":"Biophys J"},{"key":"2023041303045781000_24.4.999.8","doi-asserted-by":"publisher","DOI":"10.1038\/218438a0"},{"key":"2023041303045781000_24.4.999.9","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.91.2.669"},{"key":"2023041303045781000_24.4.999.10","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(00)81237-4"},{"key":"2023041303045781000_24.4.999.11","doi-asserted-by":"publisher","DOI":"10.1126\/science.2321027"},{"key":"2023041303045781000_24.4.999.12","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0731953100"},{"key":"2023041303045781000_24.4.999.13","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.91.10.4604"},{"key":"2023041303045781000_24.4.999.14","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(00)00031-3"},{"key":"2023041303045781000_24.4.999.15","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/0010-406X(68)90912-2","volume":"25","year":"1968","journal-title":"Comp Biochem Physiol"},{"key":"2023041303045781000_24.4.999.16","doi-asserted-by":"crossref","unstructured":"Goldberg D, Schacher S (1998) Culturing large neurons of Aplysia. 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