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Status: Bibliographieeintrag

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Verfasst von:Lischik, Colin Q. [VerfasserIn]   i
 Adelmann, Leonie [VerfasserIn]   i
 Wittbrodt, Joachim [VerfasserIn]   i
Titel:Enhanced in vivo-imaging in medaka by optimized anaesthesia, fluorescent protein selection and removal of pigmentation
Verf.angabe:Colin Q. Lischik, Leonie Adelmann, Joachim Wittbrodt
E-Jahr:2019
Jahr:March 7, 2019
Umfang:19 S.
Fussnoten:Gesehen am 25.04.2019
Titel Quelle:Enthalten in: PLOS ONE
Ort Quelle:San Francisco, California, US : PLOS, 2006
Jahr Quelle:2019
Band/Heft Quelle:14(2019,3) Artikel-Nummer e0212956, 19 Seiten
ISSN Quelle:1932-6203
Abstract:Fish are ideally suited for in vivo-imaging due to their transparency at early stages combined with a large genetic toolbox. Key challenges to further advance imaging are fluorophore selection, immobilization of the specimen and approaches to eliminate pigmentation. We addressed all three and identified the fluorophores and anaesthesia of choice by high throughput time-lapse imaging. Our results indicate that eGFP and mCherry are the best conservative choices for in vivo-fluorescence experiments, when availability of well-established antibodies and nanobodies matters. Still, mVenusNB and mGFPmut2 delivered highest absolute fluorescence intensities in vivo. Immobilization is of key importance during extended in vivo imaging. Here, traditional approaches are outperformed by mRNA injection of α-Bungarotoxin which allows a complete and reversible, transient immobilization. In combination with fully transparent juvenile and adult fish established by the targeted inactivation of both, oca2 and pnp4a via CRISPR/Cas9-mediated gene editing in medaka we could dramatically improve the state-of-the art imaging conditions in post-embryonic fish, now enabling light-sheet microscopy of the growing retina, brain, gills and inner organs in the absence of side effects caused by anaesthetic drugs or pigmentation.
DOI:doi:10.1371/journal.pone.0212956
URL:Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.

Volltext ; Verlag: https://doi.org/10.1371/journal.pone.0212956
 Volltext: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0212956
 DOI: https://doi.org/10.1371/journal.pone.0212956
Datenträger:Online-Ressource
Sprache:eng
Sach-SW:Anesthesia
 Embryos
 Fluorescence imaging
 Green fluorescent protein
 In vivo imaging
 Messenger RNA
 Pigments
 Zebrafish
K10plus-PPN:1663580499
Verknüpfungen:→ Zeitschrift

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