title: Optimal Anesthetic Regime for Motionless Three-Dimensional Image Acquisition During Longitudinal Studies of Adult Nonpigmented Zebrafish.
abstract: With many live imaging techniques, it is crucial that a deep level of anesthesia is reached and maintained throughout image acquisition without reducing zebrafish viability. This is particularly true for three-dimensional tomographic imaging modalities. Currently, the most commonly used anesthetic in the zebrafish community, MS-222 (tricaine methanesulfonate), does not allow this. We show, using a combination of both MS-222 and isoflurane, that we can significantly improve the anesthetic regime required for motionless image acquisition of live adult zebrafish. We have benchmarked this against the requirements of our novel quantitative imaging platform, compressive sensing optical projection tomography. Using nonpigmented transgenic zebrafish, we show that a combination of 175 ppm of both anesthetics improves the maintenance of deep anesthesia for prolonged periods of time and it can be used repeatedly to enable longitudinal imaging. Importantly, it does not affect the health or viability of the adult zebrafish. We also show that nonpigmented fish, with a mutated form of the gene transparent, took significantly longer to reach deep anesthesia. The anesthetic regime presented in this study should lead to significant improvements in accuracy and information achievable from imaging live adult zebrafish and in its application to longitudinal studies.
authors.affiliation: 1 Division of Medicine, University College London , London, United Kingdom .|2 CoMPLEX, University College London , London, United Kingdom .
authors.forename: Nicola
authors.initials: N
authors.lastname: Lockwood
authors.affiliation: 1 Division of Medicine, University College London , London, United Kingdom .|3 Magnus Life Sciences , London, United Kingdom .
authors.forename: Jennifer
authors.initials: J
authors.lastname: Parker
authors.affiliation: 4 UCL Fish Facility, Division of Biosciences, University College London , London, United Kingdom .
authors.forename: Carole
authors.initials: C
authors.lastname: Wilson
authors.affiliation: 1 Division of Medicine, University College London , London, United Kingdom .
authors.forename: Paul
authors.initials: P
authors.lastname: Frankel
mesh_terms: D062365:Aminobenzoates / Q000008:administration & dosage / Q000494:pharmacology*; D000758:Anesthesia / Q000662:veterinary*; D000777:Anesthetics / Q000008:administration & dosage / Q000494:pharmacology; D000818:Animals; D004359:Drug Therapy, Combination; D005786:Gene Expression Regulation; D021621:Imaging, Three-Dimensional / Q000379:methods / Q000662:veterinary*; D007530:Isoflurane / Q000008:administration & dosage / Q000494:pharmacology*; D007756:Laboratory Animal Science; D008137:Longitudinal Studies; D009154:Mutation; D010860:Pigments, Biological / Q000235:genetics / Q000378:metabolism*; D015027:Zebrafish
chemical_list: D062365:Aminobenzoates; D000777:Anesthetics; D010860:Pigments, Biological; C003636:tricaine; D007530:Isoflurane
keywords: anesthetic regime; husbandry; pigment biology; zebrafish