Surgical Removal of Spleens

2015-01-13

azim58 - Surgical Removal of Spleens

Surgical Removal of Spleens
aka splenectomy
Data:
S:\Research\CTL\Protocols\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"

* Materials
  o Autoclaved surgical instruments
  o tube with media (usually DMEM or RPMI) to put spleen into
  o avertin
  o syringe (1 mL?)
  o needle (27.5 guage needle)

* Protocols
  o Clean working area with virex and then ethanol
  o Place disposable padding down to collect any messes
  o 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)

o You may want to do a cardiac puncture before taking the spleen.
  o 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.
  o Lay the animal on its right side. Clean the incision area with
    70% ethanol.
  o With the scissors, make a left-sided skin incision, ∼2.5 cm
    long, midway between the last rib and the hip joint
  o 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
      stomach by mesentery.

o 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.
  o Cut away the mesentery and connective tissue, and remove the
    spleen (try to avoid letting any hair touch the spleen).
  o Place the spleen into the appropriate media (usually RPMI or
    DMEM) on ice.
  o Place dead mice in a bag labelled with protocol number, name (or
    advisors name), date
  o 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
Survival Surgery Removal of the.pdf

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