Physiology and Pharmacology Animal Perfusion

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					Physiology and Pharmacology:
      Animal Perfusion
       By HCS Student: Sherrell Griffin
           Mentor: Dr. Eitan Friedman
                   City College
    Physiology and Pharmacology Department
 What kind of research is done in the Physiology
       and Pharmacology department?

 Physiology is the study of living organisms and
     their parts and how they function under normal
 conditions. The main interest is the study of mechanical,
   physical and biochemical processes, which assist the
organism in keeping certain characteristics, such as body
 temperature, at a healthy level. It is important to study
how these processes communicate and how they affect the
 body. Another important purpose is research on how the
 body responds immeasurable conditions or disease. These
    studies can lead to treatment or pharmacological
Pharmacology is the science of chemical
   substances and how they interact with living
  organisms. The filed covers everything from the
structure and production of drugs to toxic effects
 and medical work. Pharmacological research will
often be used to characterize a disease or a normal
   What is the importance of this

  Physiology and pharmacology research goals are to
    explain the mechanisms that regulate normal
physiological processes, the reasons for their failure in
diseased states, and to explain the actions of drugs to
   prevent, diagnose or treat disease. To this end a
variety of molecular, biochemical, cellular and whole
organism experimental techniques are used. One such
technique, which is the high light of my presentation,
                  is animal perfusion
          Animal perfusion

    Animal perfusion is a common technique in the
physiology research laboratory. The term “perfusion”
is derived from the French verb “perfuse” meaning to
 pour over of through.” Fluid is injected into a blood
             vessel to an organ or a tissue.
        What is the purpose of
        performing a perfusion?

The purpose of performing a perfusion is to get rid of
 all the blood and replace it with par formaldehyde
fixer. It fixes tissues so it won’t degrade as fast and
              can process and save tissue.
• Add fixative to one of 500ml bottles.
• Add saline solution to the other 500 mL bottle. Open the
  fixative valve and bleed the fixative line from the bottle
  to the T-junction to eliminate air bubbles. Close the
  fixative valve, open the saline valve and bleed the saline
  line all the way to the canola. Close the saline valve.
• Anesthetize rat into a deep surgical plane using
  approved animal care protocols.
• Place animal on back in dissection tray.
• Make surgical cut along chest midline to expose rib
  cage using curved blade scalpel.
• Make lateral cuts bisecting ribs on both sides of animal;
  reflect rib cage exposing pericardium.
• Expose the heart and insert a large bore blunt needle
  (18 gauge) into the ascending aorta or left ventricle.
  This will be the entry site for the perfusate.
     Procedure (Continued)
•   Open saline stopcock and turn on peristaltic pump (if used) to
    allow saline flow.
•   Perfuse the rat with 25-50 ml of the saline solution for 10-15 min.
•   Perfuse with saline solution until the fluid is clear. Any blood
    remaining in the brain will cause staining of RBC's in subsequent
    immunohistochemical staining procedures.
•   When draining fluid is clear, open fixative stopcock and close
    saline stop cock to initiate fixation. Eyes, liver, and tongue should
    become pale as blood drains.
•   Perfuse the rat with fixative in the same manner as with saline.
•   Tissue is now ready for dissection followed by immersion in 0.1
    M phosphate buffer containing 15% sucrose for 24 hours at 4°C
    for future cryosectioning (optional).
•   Using scissors or triangular blade scalpel, snip or pierce the right
    atrium or vena cava to allow the blood, normal saline and fixative
    to leave the body during perfusion.
Scientific Implications
 Advantages                  Disadvantages
 Fixes tissues to keep it    Par formaldehyde is toxic
 longer.                     and carcinogenic meaning
                             that it is cancer causing.

•   Mentor Eitan Friedman
•   Dr. Sat
•   HCS
•   Ms. Sheveen Greene
•   City College
•   Marissa

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