Human Reproduction, Vol 13, 611-619, Copyright © 1998 by Oxford University Press
M Kraemer, C Fillion, B Martin-Pont and J Auger
This study examines the effect of varying several factors, both extrinsic
and technology-dependent, on the reconstruction of human sperm trajectories
and the derived kinematic measurements using videotapes and the Motion
Analysis Celltrak/S instrument. In semen samples from normal healthy men,
curvilinear (VCL) and straight line velocities (VSL) were found to increase
1.5-fold, and linearity (LIN) of trajectories and amplitude of lateral head
displacement (ALH) increased 1.25-fold when the temperature of analysis was
raised from 24 to 37 degrees C. Only VCL and VSL were found to increase
significantly between 24 and 37 degrees C for sperm samples selected by
Percoll gradient and incubated in a capacitating medium. An analysis
chamber of 20 microm depth was found to be adequate for seminal sperm
samples while for Percoll-selected sperm samples the analysis in a 50
microm depth provided the highest proportions of spermatozoa with the
highest VCL and the largest ALH. The grey level detection threshold
required careful adjustment: using a threshold lower than the optimal
threshold produced spurious sperm trajectories for seminal sperm samples
and rejected some trajectories for Percoll-selected sperm samples.
Definition of the appropriate frame rate and maximum burst speed was
critical for valid trajectory reconstruction and therefore adequate derived
kinematic measurements. Optimal values of these parameters were found to be
30 Hz and 400 microm/s for seminal spermatozoa and 60 Hz and 700 microm/s
for selected spermatozoa. The optimal values of 'ALH path-smoothing factor'
used to calculate average path and ALH were 5-10 points for seminal
spermatozoa analysed at 30 Hz and 15-20 points for selected spermatozoa
analysed at 60 Hz. We propose a set of standard conditions for reliable
kinematic analysis of human spermatozoa using the Celltrak/S system.
ARTICLES
Factors influencing human sperm kinematic measurements by the Celltrak computer-assisted sperm analysis system
Laboratoire de Biologie de la Reproduction, Hopital Jean Verdier, Bondy, France.
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