Current protocols in immunology by John E Coligan; et al

By John E Coligan; et al

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0 150 aAdapted from Tuffery, 1987. globin levels should be monitored to preclude anemia when frequent blood samples are taken. Use of anesthetics is recommended for some of the protocols. Anesthetics may alter some hematological and biochemical parameters, but their use is indicated to prevent undue stress. When repeated venipuncture is required over extended periods, surgical placement of indwelling catheters (arterial or venous) is recommended. Descriptions of such techniques are reviewed by Gay (1986).

The inexperienced investigator should take the trouble to gain this skill in several practice sessions with phosphate-buffered saline (PBS) as the injectate. 3) Heat lamp or beaker containing warm water Gauze sponge or swab 1. Fill syringe with injectate and remove air bubbles. Removal of air bubbles is critical to avoid air embolism. 2. Place mouse in a restrainer. 3. Warm the tail with a heat lamp or by immersing in warm water to dilate vessels. 4. Swab the tail with 70% ethanol on a gauze sponge or swab.

Follow steps 1 to 5 of the basic protocol for splenectomy. 2. Minimize bleeding by placing a small loop of suture across the center of the spleen, leaving the hilar vessels intact. Gently tighten the suture and tie it. Tightening too quickly will split the spleen, especially if there is moderate splenomegaly prior to surgery. 3. Cut the lower part of the spleen with scissors and remove it. 4. Close the animal as in the basic protocol for splenectomy, steps 8 to 11. BASIC PROTOCOL NEONATAL THYMECTOMY Neonatal thymectomy is used most often to remove mature T cells without removing T cell precursors.

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