2. Additional materials needed: Lab Top Incubator SpermPrep™ Media Syringe with needle IUI Catheter Pipette

3. Remove the Multi- ZSC™ from pouch and keep warm (37 ⁰ C) throughout the procedure. Maintaining physiological temperature is very important to insure maximal yield of healthy, motile sperm. Use an incubator for incubation.

4. Remove top stopper and place 2.0mL of liquefied semen inside the cone on bottom with pipette until completely filled. Avoid introducing air bubbles during placement of semen. Also, the use of non- liquefied, viscous specimens can prevent the recovery of adequate numbers of sperm.

5. Using a tuberculin syringe with needle, place 1.0mL of media (SpermPrep ™ ) in the area around the cone containing the semen, and make sure that the top of the cone containing the semen is completely covered. Continue to fill the various compartments of the Multi- ZSC ™ one by one with approximately 1.0mL of media each. Cap the Multi- ZSC ™ with top stopper.

6. Place the Multi- ZSC ™ in an incubator at 37 ⁰ C and incubate for 60 minutes. For ISCI or other ART procedures, you may wish to shorten or prolong the incubation time, according to the numbers of sperm you wish to recover. If specimen is deficient, you may incubate longer.

7. Remove the Multi-ZSC ™ from incubator. Using syringe with needle, remove all media placed in Multi-ZSC ™ by compartment, by placing needle on the outside of the bottom of each cone and aspirating until all media containing the healthy sperm is removed from each compartment (approximately 1.0mL). Label and keep the media from each compartment separate.

8. To use the prepared semen, simply remove the needle from the syringe that contains the recovered spermatozoa, attach the IUI catheter, and perform the IUI procedure. You may used the recovered sperm in media as is, or you may choose to pool and concentrate the recovered sperm to desirable levels via centrifugation prior to performing an ART procedure.

9. To order your ZSC Kits, LabTop incubator, SpermPrep Media, and IUI Catheters, please call us at 1-800-998-4567 or1-859-278-6806 You may also e-mail us you order at sales@zdlinc.com Or visit our website www.zdlinc.net