Surgical+Removal+of+Spleens

aka splenectomy Data: S:\Research\CTL\Protocols\surgical removal of spleens
 * Surgical Removal of Spleens**

see also a protocols from Deborah Hansen "F:\kurt\storage\CIM Research Folder\DR\2013\5-8-13\spleen and blood collection protocols\20130507 final bleeds + spleen.xlsx" "F:\kurt\storage\CIM Research Folder\DR\2013\5-8-13\spleen and blood collection protocols\20130507 Splenocyte Preparation DTH.docx"

>>> stomach by mesentery.
 * Materials
 * Autoclaved surgical instruments
 * tube with media (usually DMEM or RPMI) to put spleen into
 * avertin
 * syringe (1 mL?)
 * needle (27.5 guage needle)
 * Protocols
 * Clean working area with virex and then ethanol
 * Place disposable padding down to collect any messes
 * Inject mouse with anaesthesia. Wait several minutes for anaesthesia to take effect. The mouse should be unresponsive when the hind feet are squeezed. If the mouse is still responsive, inject more anaesthesia.
 * We typically use avertin (300 uL of 0.05 mg/mL diluted in H20?) injected intraperitoneally (i.p.) with 27.5 guage needle (see intraperitoneal mouse injection)
 * You may want to do a cardiac puncture before taking the spleen.
 * Before any surgical procedures snap neck by holding the tail, pushing down on the neck with a plunger, and pushing the head away from the tail.
 * Lay the animal on its right side. Clean the incision area with 70% ethanol.
 * With the scissors, make a left-sided skin incision, ∼2.5 cm long, midway between the last rib and the hip joint
 * Loosen any connective tissue under the skin using the blunt end of the forceps.
 * The spleen is easily seen through the peritoneum attached to the greater curvature of the
 * Make a 1- to 2-cm incision in the peritoneal wall and gently pull the spleen onto the exterior surface of the peritoneum. A small artery, not always visible, is attached to the hilum of the spleen, closest to the stomach.
 * Cut away the mesentery and connective tissue, and remove the spleen (try to avoid letting any hair touch the spleen).
 * Place the spleen into the appropriate media (usually RPMI or DMEM) on ice.
 * Place dead mice in a bag labelled with protocol number, name (or advisors name), date
 * When these mice are placed in the freezer in the other room the weight may need to be estimated (about 17 g per mouse)

Associated Literature Survival Surgery of Spleen: http://www.currentprotocols.com/protocol/im0110