MOTILITY AND FERTILITY

CORRELATION
 SEMEN ANALYSIS

Subjective Biochemical or Biological tests Objective

Estimation of
Metabolic activity
Gross Motility Respiratory activity Zona free Hamster ova Time-lapse photo micrography
or Mass penetration assay or Sperm
activity (0-5) Penetration assay
Progressive MBRT Cervical mucus penetration frame by frame video micrography
Motility (0- test
100%)
Fructolysis activity Spectophotometry

CASA
 COMPUTER ASSISTED SPERM ANALYSIS

• MICROSCOPE – DIGITAL CAMERA – COMPUTER WITH SOFTWARE ALGORITHM

• SERIES OF SUCCESSIVE IMAGES OF MOTILE SPERMATOZOA WITHIN A STATIC
FIELD OF VIEW
• SPERM CONCENTRATION
• MORPHOLOGY
• VIABILITY
• AND INDEX OF DNA FRAGMENTATION OF FROZEN THAWED SPERM
 CASA

• POSITIVE RELATIONSHIP
• CASA PROVIDES NUMBERS OF OBJECTIVE MEASURES OF SPERM MOTION CHARACTERISTICS
TAKEN FROM TRACKS OF LARGE NUMBER OF SPERM
• OPTIMUM SPERM CONCENTRATION – 20 X 106/ML
• DURATION OF ANALYSIS – SHOULD NOT EXCEED 5 MINS
• VOLUME – 4 TO 10 ΜL
• MICROCELL CHAMBER – 7 ΜL
• MAKLER CHAMBER – 4 ΜL
• LEJA – 4 ΜL

• SERGE –SILBERBERG EFFECT

 CASA PARAMETERS

• CURVILINEAR VELOCITY – VCL

• AVERAGE PATH VELOCITY – VAP
• STRAIGHT LINE VELOCITY – VSL
• STRAIGHTNESS – STR (VSL/VAP)
• LINEARITY – LIN (VSL/VCL)
• WOBBLE – WOB (VAP/VCL)
• BEAT CROSS FREQUENCY (HZ)
 PARAMETERS
• CURVILINEAR VELOCITY (VCL ΜM/SEC) – SUM OF THE DISTANCES BETWEEN EACH MEASURED SPERM POSITION DIVIDED
BY THE ANALYSIS TIME.

• STRAIGHT LINE VELOCITY (VSL ΜM/SEC) – STRAIGHT LINE DISTANCE BETWEEN THE FIRST AND LAST SPERM POSITION
DIVIDED BY THE ANALYSIS TIME.

• AVERAGE PATH VELOCITY (VAP ΜM/SEC) – THE TIME AVERAGED VELOCITY OF A SPERM HEAD ALONG ITS AVERAGE
PATH

• STRAIGHTNESS (STR = VSL/VAP) - MEASURE OF LINEARITY OF THE AVERAGE PATH

• LINEARITY OF FORWARD PROGRESSION (LIN = VSL/VCL) - IS THE LINEARITY OF THE CURVILINEAR PATH

• WOBBLE (WOB = VAP/VCL) - MEASURE OF OSCILLATION OF THE ACTUAL PATH ABOUT THE AVERAGE PATH

• AMPLITUDE OF LATERAL HEAD DISPLACEMENT (ALH ΜM) - IS THE AVG. DISTANCE OF THE SPERM HEAD FROM THE AVG.
SPERM SWIMMING PATH
 CASA PARAMETERS
• TOTAL MOTILITY – TMOT
• PROGRESSIVE MOTILITY – PMOT
• AMPLITUDE OF LATERAL HEAD DISPLACEMENT – ALH
• TWICE THE MAXIMUM DISPLACEMENT OF SPERM HEAD FROM ITS FITTED MOVING AXIS IN A TRACK
SEGMENT (ΜM) – DIRECTLY RELATED TO THE TAIL BENDING (AT PROXIMAL REGION OF TAIL)
• LARGER ALH – STRONGER BENDING

• BEAT CROSS FREQUENCY – BCF

• FREQUENCY THAT THE SPERM HEAD MOVES ACROSS THE MIDDLE PLANE OF THE STRAIGHTENED
TRAJECTORY (UNIT : HZ)
• SUBLINERALY PROPORTIONAL TO THE BEATING FREQUENCY OF THE SPERM TAIL
• ROUGHLY DOUBLE THE FREQUENCY OF HEAD WOBBLING

• LOW VAP CUT OFF – LVV

• MEDIUM VAP CUT OFF - MVV
 WHO STANDARD 2010 (HUMAN)

• % OF PROGRESSIVE MOTILITY BY VAP WHICH EXCEEDS MVV AND THE MINIMAL

VALUE OF STR
• CATEGORIES OF SPERMATOZOA
• FAST VAP > MVV
• MEDIUM FAST LVV < VAP << MVV
• SLOW VAP < LVV
• IMMOTILE -
 CORRELATION

• PARAMETERS WITH HIGH CORRELATION WITH BULL FERTILITY(FARREL ET AL.

1998)
• ALH, BCF, LIN, VAP, VSL (R2 = 0.98)
• BCF, LIN, VSL, VCL, STR (R2 = 0.97)
• BCF, LIN, VAP, VSL, STR (R2 = 0.97)
• BCF, LIN, VAP, VSL, VCL (R2 = 0.97)

• SINGLE PARAMETER R2 VALUES ARE LOWER

 ESTIMATE FOR LOW FIELD FERTILITY

• LOW FIELD FERTILITY

• CSEH ET AL. 2004

• VCL – 22.51 TO 79.5 ΜICRO METER/S

• VSL – 11.30 TO 35.71 ΜICRO METER/S
• VAP – 12.67 TO 34.45 ΜICRO METER/S
 SPERM SUB POPULATIONS

• FIRST
• RELATIVELY LOW VELOCITY BUT HIGH PMOT (HIGH LIN, STR, WOB, BCF & LOW ALH)
• SECOND
• HIGH ACTIVITY BUT WITHOUT PMOT (HIGH VCL & ALH LOW LIN & STR)
• THIRD
• VERY LOW MOTILITY & WITHOUT PMOT (LOW VCL, VSL, VAP, ALH, BCF, LIN, STR, WOB)
• FOURTH
• FAST AND PMOT HIGH VALUE OF BOTH (I) VCL, VSL, VAP, BCF(II) LIN, STR, WOB, ALH

• BEFORE CRYOPRESERVATION
• FAST AND PMOT
• CORRELATES WITH THE LONGEVITY OF SPERM (NUNEZ-MARTINEZ ET AL. 2006)
 TIME LAPSE PHOTOMICROGRAPHY

• CLOSE PROXIMITY
• SPERM PATH INTERSECT
• JPDAF – JOINT PROBABILISTIC DATA ASSOCIATION FILTER
• AIR TRAFFIC CONTROL SYSTEMS
• INDEPENDENT KALMEN FILTERS – ESTIMATE THE POSITION AND VELOCITY OF
EACH SPERM TRACKED
• ENABLES ACCURATE TRACKING OF SPERM IN UNDILUTED SPECIMEN
THANK YOU