{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T15:16:12Z","timestamp":1772118972032,"version":"3.50.1"},"reference-count":35,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2018,5,27]],"date-time":"2018-05-27T00:00:00Z","timestamp":1527379200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01 AI120902"],"award-info":[{"award-number":["R01 AI120902"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["U01 EB021238"],"award-info":[{"award-number":["U01 EB021238"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000011","name":"Howard Hughes Medical Institute","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000011","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Small Methods"],"published-print":{"date-parts":[[2018,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Single\u2010molecule fluorescence in situ hybridization (smFISH) provides direct access to the spatial relationship between nucleic acids and specific subcellular locations. The ability to precisely localize a messenger RNA can reveal key information about its regulation. Although smFISH is well established in cell culture or thin sections, the utility of smFISH is hindered in thick tissue sections due to the poor probe penetration of fixed tissue, the inaccessibility of target mRNAs for probe hybridization, high background fluorescence, spherical aberration along the optical axis, and the lack of methods for image segmentation of organelles. Studying mRNA localization in 50 \u00b5m thick\n                    <jats:italic>Drosophila<\/jats:italic>\n                    larval muscle sections, these obstacles are overcome using sample\u2010specific optimization of smFISH, particle identification based on maximum likelihood testing, and 3D multiple\u2010organelle segmentation. The latter allows independent thresholds to be assigned to different regions of interest within an image stack. This approach therefore facilitates accurate measurement of mRNA location in thick tissues.\n                  <\/jats:p>","DOI":"10.1002\/smtd.201700324","type":"journal-article","created":{"date-parts":[[2018,5,28]],"date-time":"2018-05-28T02:40:35Z","timestamp":1527475235000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Advanced 3D Analysis and Optimization of Single\u2010Molecule FISH in\n                    <i>Drosophila<\/i>\n                    Muscle"],"prefix":"10.1002","volume":"2","author":[{"given":"Akiko","family":"Noma","sequence":"first","affiliation":[{"name":"RNA Therapeutics Institute University of Massachusetts Medical School  Worcester MA 01605 USA"}]},{"given":"Carlas S.","family":"Smith","sequence":"additional","affiliation":[{"name":"RNA Therapeutics Institute University of Massachusetts Medical School  Worcester MA 01605 USA"}]},{"given":"Maximiliaan","family":"Huisman","sequence":"additional","affiliation":[{"name":"RNA Therapeutics Institute University of Massachusetts Medical School  Worcester MA 01605 USA"}]},{"given":"Robert M.","family":"Martin","sequence":"additional","affiliation":[{"name":"Instituto de Medicina Molecular Faculdade de Medicina Universidade de Lisboa  1600\u2010276 Lisboa Portugal"}]},{"given":"Melissa J.","family":"Moore","sequence":"additional","affiliation":[{"name":"RNA Therapeutics Institute University of Massachusetts Medical School  Worcester MA 01605 USA"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9067-804X","authenticated-orcid":false,"given":"David","family":"Grunwald","sequence":"additional","affiliation":[{"name":"RNA Therapeutics Institute University of Massachusetts Medical School  Worcester MA 01605 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