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v i r a l   i m m u n i t y   &   p a t h o g e n e s i s   g r o u p

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OUTLINE

Experimental splenectomy in the mouse is used to study the role of the spleen in platelet destruction..

PROTOCOL

General anesthesia
  1. inject i.p. 250l/mouse of the ANESTHETIC mixture < wait for 2-10min for the anesthetic effect>
  2. for verify the anesthetic effect check plantar reflexes (animal must be unresponsive to squeezing hind feet)

Preparation of the operation area
1.    wash the surgical and adjacent area with C 2 H5OH-70%
2.    raze the fur with the razor
3.    wash the surgical area with C2 H 5 OH-70%

Operation
  1. At the left hypochondrium make a 1.5-2cm "skin, muscles and peritoneum" incision with surgical scissors
  2. Gently trap and push the spleen free of adjacent tissue, separate its leg with blunt-ended scissors or forceps
  3. By a single knot tie off the artery with a 3-0 plane CATGUT by looping the suture through the mesentary (the artery enters the spleen at the superior polus). If necessary by a single knot tie off the efferent venule (exits the spleen at the inferior polus)
  4. Cut away the connective tissue and remove the spleen
  5. Apply on the open wound 1 pinch of Astrexene (Chlorhexidin HCl) (5mg/g)
  6. Apply on the peritoneum and muscles the U-like suture with plane CATGUT (3-0)
  7. Apply on the skin the U-like suture with ETHICON MERSILENE (3-0)
Wakening
1.   inject 250ul/mouse of the "WAKENING MIXTURE"

Post operation period
1.    after the operation took place let the animals to rest for at least 7 days in a cage with the filter
2.    observe animals every day

SOLUTIONS
  1. ANESTHETIC MIXTURE=mix 1ml of Domitor (Medetomidin HCl) (1mg/ml) , 1ml of Imalgene (Ketaminum HCl) (50mg/ml) and 8ml of PBS <all steril>.
  2. "WAKENING MIXTURE"=mix 1ml of ANTISEDAN (Atipamezole HCl) (5mg/ml) and 9ml of PBS <all sterile!> 
ADDITIONAL INFO

Survey the mice for some time after this operation.


created by Andrei Musaji          ::          updated : 2004