Euthanasia: All protocols involving animals in this work were approved by Institutional Animal Care and Use Committees (IACUC) of USC.
Specimens were collected off the Southern California coast by Chuck Winkler* and transported to USC the same day. Euthanasia was accomplished by immersion in a 1:3 mixture of isotonic magnesium chloride (75g/L MgCl2 in D.I.) in artificial sea water. Specimen was maintained in this solution for at least 10 minutes after cessation of breathing, spontaneous behavior and motion of any kind before removing (1, 2). Specimen was transferred to 4% paraformaldehyde solution with stirring at 10ºC for several days for fixation.
MRI: Sample was soaked in gadolinium based contrast agent for 3 days. Sample was immersed in Galden (TMC Industries, MN) while being scanned.
CT: Sample was soaked in Lugol’s iodine solution (VWR, GA) for 3-5 days.
*Chuck Winkler, PharmD
Aquatic Research Consultants
San Pedro, CA 90732
Website: Fishes4Study
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- Pugliese C, Mazza R, Andrews PLR, Cerra MC, Fiorito G, Gattuso A. Effect of Different Formulations of Magnesium Chloride Used As Anesthetic Agents on the Performance of the Isolated Heart of Octopus vulgaris. Frontiers in Physiology. 2016;7(610). doi: 10.3389/fphys.2016.00610.