The video describes a sampling method for peripheral blood collection from retro-orbital sinus of a mouse and further isolation of mononuclear cells from blood with density gradient centrifugation.
Protocol
1. Retro orbital blood collection Perform retro orbital blood collection just before euthanasia under sterile conditions in a laminar flow hood and under a heating lamp to prevent hypothermia. For anesthesia, use ketamine at 150 mg/kg and xylazine at 10 mg/kg. Prepare the anesthetic solution by diluting 1.5 ml of ketamine and 0.5 ml of xylazine in 18 ml of PBS solution. Insert a capillary tube into the medial canthus of the eye. Blood will rise from the orbital sinus into …
Retro-Orbital Blood Sampling: A Method for Isolating Mononuclear Cells from the Retro-Orbital Sinus of a Mouse. J. Vis. Exp. (Pending Publication), e20263, doi: (2023).