{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T13:51:46Z","timestamp":1791381106560,"version":"4.3.3"},"reference-count":21,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T00:00:00Z","timestamp":1791331200000},"content-version":"vor","delay-in-days":6,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"},{"start":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T00:00:00Z","timestamp":1790812800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100010926","name":"Illinois State University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100010926","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Spreading depolarization (SD) is a ubiquitous neural phenomenon implicated in mammalian neurological disorders including cerebral ischemia, brain death, stroke, brain injury, and migraine, as well as in insect chill coma. During SD, a breakdown of ion homeostasis and a slowly spreading wave of neural depolarization leads to processing deficits, such as sensory disturbances and paralysis. Our protocols introduce high\u2010throughput techniques using wide\u2010field fluorescence brain imaging in\n                    <jats:italic>Drosophila melanogaster<\/jats:italic>\n                    larvae and adults to detect and quantify the spatial and temporal dynamics of SD and its initiation points. Two SD initiation techniques, rapid cooling and pharmacological manipulation, are detailed. The larval protocols describe how to elicit, detect, and quantify SD in up to 16 animals simultaneously in either intact or dissected preparations. The adult SD protocols describe the detailed steps to elicit, detect, and quantify SD in either fully intact animals or after microdissection that enables high resolution imaging of the brain. Detection, quantification, and analysis techniques are described in a separate protocol. Our protocols establish the fly as a high\u2010throughput model for studying conserved cellular and circuit\u2010level mechanisms of SD initiation and propagation. \u00a9 2026 The Author(s).\n                    <jats:italic>Current Protocols<\/jats:italic>\n                    published by Wiley Periodicals LLC.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : High\u2010throughput SD detection in intact\n                    <jats:italic>Drosophila<\/jats:italic>\n                    larvae with fluorescent calcium reporters\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Alternate Protocol 1<\/jats:bold>\n                    : High\u2010resolution SD imaging in fillet larvae with fluorescent calcium reporters\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 1<\/jats:bold>\n                    : SD imaging in larvae with fluorescent membrane voltage reporters\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : High\u2010throughput SD detection in adult\n                    <jats:italic>Drosophila<\/jats:italic>\n                    with fluorescent calcium reporters\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Alternate Protocol 2<\/jats:bold>\n                    : High\u2010resolution SD imaging in adult\n                    <jats:italic>Drosophila<\/jats:italic>\n                    with fluorescent calcium reporters\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 2<\/jats:bold>\n                    : SD imaging in adults with fluorescent membrane voltage reporters\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 3<\/jats:bold>\n                    : High\u2010throughput SD detection analysis in\n                    <jats:italic>Drosophila<\/jats:italic>\n                    with fluorescent calcium reporters\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Alternate Protocol 3<\/jats:bold>\n                    : High\u2010resolution SD imaging analysis in\n                    <jats:italic>Drosophila<\/jats:italic>\n                    with fluorescent calcium reporters\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 3<\/jats:bold>\n                    : SD detection and spread analysis in\n                    <jats:italic>Drosophila<\/jats:italic>\n                    with fluorescent membrane voltage reporters\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70475","type":"journal-article","created":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T12:57:47Z","timestamp":1791377867000},"source":"Crossref","is-referenced-by-count":0,"title":["Detection and Quantification of Spreading Depolarization in\n                    <i>Drosophila melanogaster<\/i>\n                    Using High\u2010Throughput, Wide\u2010Field Brain Imaging"],"prefix":"10.1002","volume":"6","author":[{"given":"Wolfgang","family":"Stein","sequence":"first","affiliation":[{"name":"School of Biological Sciences Illinois State University  Normal Illinois"},{"name":"These authors contributed equally to this work"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Allison L.","family":"Harris","sequence":"additional","affiliation":[{"name":"Department of Physics Illinois State University  Normal Illinois"},{"name":"These authors contributed equally to this work"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,7]]},"reference":[{"key":"e_1_2_15_2_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.2006732"},{"key":"e_1_2_15_3_1","doi-asserted-by":"publisher","DOI":"10.1242\/jeb.246313"},{"key":"e_1_2_15_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jinsphys.2012.09.006"},{"key":"e_1_2_15_5_1","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00027.2014"},{"key":"e_1_2_15_6_1","doi-asserted-by":"publisher","DOI":"10.1002\/ana.25147"},{"key":"e_1_2_15_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2026.116389"},{"key":"e_1_2_15_8_1","doi-asserted-by":"crossref","unstructured":"Harris A. &Stein W.(2026). 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