UWM ARC Sentinel Serology Sample Collection and Submission Procedure for Rats

Rat Serum Sample SOPpdf format, Adobe Acrobat Required(for printing)

Serum Collection, Preparation and Shipping:

  1. Identify rat to be tested and anesthetize via chamber induction and nose cone maintenance.
  2. Draw 300-600 microliters of blood from lateral tail vein or other suitable site, using 1ml tuberculin syringe or 1ml syringe with 25-26 Ga hypodermic needle. Draw blood slowly to avoid hemolysis in the narrow-gauge needle.
  3. Gently eject the whole blood into a 1.6ml microcentrifuge tube. (Cap the needle using a one-handed capping procedure, remove and discard the needle in a sharps container, and eject the blood into the microcentrifuge tube.)
  4. Place the microcentrifuge tube into the centrifuge, along with a counter-balancing microcentrifuge tube filled with a small amount of water. Centrifuge the blood at 5000 rpm for approximately 10 minutes. Rebalance if necessary, if excessive vibration is present.
  5. Using a 100µl pipette, pipette the serum component from the centrifuge tube into a clean 0.6 or 1.6 ml microcentrifuge tube.
  6. Optional: remix the blood in the original tube and repeat steps 4 and 5 to obtain additional serum.
  7. Optional: If a reddish tinge is observed in the serum tube, centrifuge this tube at 5000 rpm for 10 minutes, and pipette off the clear serum into a second (clean) microcentrifuge tube.
  8. If less than 100µl of serum is obtained, dilute the serum at a 1:5 dilution (1 part serum to 4 parts physiologic saline or phosphate-buffered saline).
  9. Label the microtube of serum with a permanent marker and freeze at -20°C overnight. At this time, consider also freezing a block of ice with an embedded styrofoam insert for shipping. (Do not use dry ice to ship.)
  10. Complete the RADIL Lab Submission form; don’t forget to indicate the serum dilution.
  11. Place the frozen microtube and block of ice (or equivalent cooling block) in a sealed bag, and package for shipping to MU-RADIL.
  12. Label the package (label found by fax machine in Eng. 270) and send via FedEx Overnight Service. It is ok to send it out anytime during the day for next day delivery. There is a Fed Ex on Brady Street. Use the state visa to pay for this.

Parasitology:

Fecal Float:

  1. Collect feces from the cage and place in a test tube. Mix with Zn Sulfate solution—about half way and mash feces.
  2. Pour off the fluid into a clean test tube through gauze to strain it.
  3. Add enough Zn sulfate to fill the test tube to the top and put a coverslip on the top.
  4. Let it sit for 2 hours.
  5. Put coverslip on a slide and view under scope at 10x for eggs.

Cellophane Tape Test:

  1. Use clear tape and rub area around the rats anus.
  2. Put tape, sticky side down, on a slide.
  3. View under scope at 10x for eggs.

Updated June 18, 2007 by SAK