1. Spermatology 2006 1 IVOS & CEROS Computer Assisted Sperm Analysis Spermatology 2006

2. 2 What is CASA?  An automated system designed to: Visualize and digitize successive images of sperm Process and analyze the images Provide precise and meaningful information on sperm kinematics and counts  Combination of hardware and software Hardware: optics, illuminator, digitizer, computer Software: modules for motility, morphology, and viability analysis

3. Spermatology 2006 3 2 Current CASA Models from HT  IVOS Sperm Analyzer Built-in optical system Automated, heated stage Fastest analysis: 6 fields in 6 seconds  CEROS Sperm Analyzer Complete system Includes negative phase contrast microscope

4. Spermatology 2006 4 NEW!! Modular CEROS  Choose only the options you need  Cost-effective  Use your own negative or positive phase contrast microscope  Same proven and trusted analysis algorithms

5. Spermatology 2006 5 Multi-species Compatibility  Human  Non-human primates  Domestic / companion animals  Production animals  Endangered species  Laboratory rodents  Sea mammals and fish

6. Spermatology 2006 6 Customized CEROS for Fish Sperm  “Cooled” stage for optimum maintenance of fish sperm motility  Applications include: Monitoring sperm motility at hatcheries Testing extension media and spermiation stimulant Monitoring environmental pollutants Reproductive toxicology  Fish, sea urchin, etc.

7. Spermatology 2006 7 Negative Phase Contrast - Standard 10x Negative Phase20x Negative Phase Provides fastest analysis Provides superb visualization

8. Spermatology 2006 8 Getting Ready for Analysis (IVOS)  Load sample in fixed or known depth chamber (e.g. Leja Chamber, Microcell, Makler)  Place filled chamber on stage  Select multiple analysis fields  Press Start Scan

9. Spermatology 2006 9 Sperm Analysis  Captures multiple “snapshots” extremely quickly (e.g., 30 images in 0.5 second).  Tracks the motile cells from snapshot to snapshot  Static sperm are identified based on motile cell means (size, contrast, intensity)  Computer calculates results

10. Spermatology 2006 10 Results  Motility  Progressive Motility  Concentrations  Counts  Velocity - 3 types  Motion parameters

11. Spermatology 2006 11 Playback for Quality Control  Motile sperm marked with green track  Progressive sperm marked with blue track  Slow sperm marked with purple track  Static sperm marked with red dot

12. Spermatology 2006 12 Motile Playback Sample

13. Spermatology 2006 13 Research Options  Obtain data on individual tracks (Edit Tracks)  Study sub-populations of sperm (Sort)

14. Spermatology 2006 14 Data Management  Clinical Filing Custom designed reports Store reports to disk  HDATA ASCII Export Export data in database / spreadsheet compatible format Summary and individual track data

15. Spermatology 2006 15 Digital Image Storage  Ability to save all fields analyzed  Recall and reanalyze fields  Sequential field analysis – all fields from original analysis automatically analyzed in order  Share images with other IVOS/CEROS users  Create QC images for training purposes

16. Spermatology 2006 16 Morphology Options (Automated)  Dimensions Morphology Human sperm only Analysis based on Dr. Kruger’s Strict Criteria Diff-Quik and PAP stains  Metrix Morphology Multi-species Analysis based on user’s definition of normal Diff-Quik, PAP and other high contrast stains

17. Spermatology 2006 17 Real-Time Morphology (non-automated)  Subjective (visual classification)  Fast – “REAL-TIME”  Use non-stained, immobilized sperm, or standard stained smear  Results combined with motility results  Morph-merge option

18. Spermatology 2006 18 IDENT Fluorescence (IVOS Only)  Static and motile sperm identification for high- detritus samples  Sperm stained with Hoechst 33342 stain  All sperm fluoresce under UV light  Accurate counts to within 1%

19. Spermatology 2006 19 VIADENT Fluorescence (IVOS Only)  Sperm stained with Hoechst 33258 stain  Only NON-VIABLE sperm fluoresce under UV illumination  Counting Motile/Total counts under standard illumination Viable count under fluorescent illumination  Only system to perform motility and viability analysis on the same, live sample

20. Spermatology 2006 20 Benefits of CASA  Objectivity: Removes technician bias  Accuracy: Best method for determining percent motility, percent progressive motility, and velocity parameters  Precision: Repeatable results  Sensitivity: Detects even the slightest changes in sperm motion  Reliability: Does not get “tired eyes” – can analyze samples all day without sacrificing accuracy

21. Spermatology 2006 21 Hamilton Thorne Contact Information  Phone: 978-921-2050, 800-323-0503  Fax: 978-921-0250  E-mail: info@hamiltonthorne.com  Web: www.hamiltonthorne.comwww.hamiltonthorne.com  100 Cummings Center, Suite 465E Beverly, MA 01915 USA