THREE SIMPLE STEPS FOR PROCESSING SEMEN VIA SPERMPREP™II FILTER (SP-II) SpermPrep™II IT IS AS SIMPLE AS A-B-C! All specimens should be “washed” prior to filtration except post-thaw specimens. Before using the SpermPrep™II (SP-II) filter, we suggest that you get acquainted with its various parts by considering the diagram shown on the this page and comparing it with the actual filter itself. Complete familiarity with the device (SP-II) will be quite helpful in properly employing and acquiring the best results during use.

THREE SIMPLE STEPS FOR PROCESSING SEMEN VIA SPERMPREP™II FILTER (SP-II) SpermPrep™II IT IS AS SIMPLE AS A-B-C! A. HYDRATION Remove top stopper and place 6.0 mL of warm media (37⁰C) in barrel of SP-II. Remove top disc and mix the beads by bringing them into suspension via a pipette. Allow to set in a warm environment (warm plate or incubator) for 10 minutes. Remove bottom closure and allow the media to drip for 2 minutes, then close it and discard filtrate. Add more media to bring to 4-5 mL level. The SP-II is now ready for use.

THREE SIMPLE STEPS FOR PROCESSING SEMEN VIA SPERMPREP™II FILTER (SP-II) SpermPrep™II IT IS AS SIMPLE AS A-B-C! B. FILTRATION Place aliquot of washed, reconstituted semen, which should contain a maximum of 250 x 10⁶ total sperm in barrel of SP-II, mix gently with existing media (from Step A), open the SP-II by removing the bottom closure and filter for 7-10 minutes.

THREE SIMPLE STEPS FOR PROCESSING SEMEN VIA SPERMPREP™II FILTER (SP-II) SpermPrep™II IT IS AS SIMPLE AS A-B-C! C. RECONSTITUTION OF FILTRATE Centrifuge the filtrate (300 G for 7 minutes) and suspend the whole sperm pellet in volume of choice for intrauterine AI or other assisted reproductive technique (ART).

TROUBLESHOOTING SpermPrep™II 1.Slow filtration (slow recovery of filtrate) You may not be maintaining adequate media level You may be overloading the filter with sperm or excessive debris 2.Recovery of sperm with excessive debris in filtrate You may be overloading the filter with sperm and debris 3.Recovery of Sephadex beads in filtrate* You may be improperly hydrating and priming SP-II filter NOTE: Sephadex beads are made out of sugar molecules and are not toxic if accidentally placed intrauterinely when performing IUI