view - Department of Reproduction, Obstetrics and Herd Health
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CHAPTER 3.2<br />
88<br />
In addition, it has been demonstrated that chamber depth influences human sperm motility<br />
parameters (Le Lannou et al., 1992). Also in veterinary medicine, the effect <strong>of</strong> the chamber type has<br />
been analyzed for a few chambers (Contri et al., 2010; Iguer-ouada <strong>and</strong> Verstegen, 2001; Lenz et al.,<br />
2011). The Makler chamber was associated with higher motility outcomes compared to the Leja <strong>and</strong><br />
Cell-vu chambers (Contri et al., 2010; Iguer-ouada <strong>and</strong> Verstegen, 2001; Lenz et al., 2011), but was<br />
comparable to the WHO slide (Lenz et al., 2011). In literature, the depth <strong>of</strong> the sperm suspension for<br />
motility analysis is variable <strong>and</strong> not seldom left unmentioned which is in sharp contrast to human<br />
<strong>and</strong>rology where the WHO laboratory manual for the examination <strong>and</strong> processing <strong>of</strong> human semen<br />
(WHO, 2010) clearly defines how a slide for sperm motility analysis should be prepared. This is<br />
particularly important since it is known that a chamber depth <strong>of</strong> less than 20 µm constrains the<br />
rotational movement <strong>of</strong> human spermatozoa (Kraemer et al., 1998; Le Lannou et al., 1992). Ishijima<br />
<strong>and</strong> co-workers (1986) showed that the flagellar beating <strong>of</strong> a spermatozoa was three dimensional<br />
rather than planar <strong>and</strong> results in a sperm rotation on a helical trajectory. As such, a shallow chamber<br />
<strong>of</strong> 10 µm depth prevents the development <strong>of</strong> highly curved flagellar beats; as such the trajectory<br />
remains straight <strong>and</strong> the rate <strong>of</strong> progressive hyperactivated motions increases (Le Lannou et al.,<br />
1992). The differences in width <strong>of</strong> sperm heads between human <strong>and</strong> animal species (Table 1) might<br />
affect motion characteristics to a different extent.<br />
Table 1. Dimensions <strong>of</strong> the sperm head <strong>of</strong> different mammalian species.<br />
Human 1<br />
Equine 2<br />
Bovine 3<br />
Canine 4<br />
Porcine 5<br />
Length (µm) 4.1 5.49 8.67 6.65 8.12<br />
Width (µm) 2.8 2.65 4.55 3.88 4.07<br />
1 WHO laboratory manual for the examination <strong>and</strong> processing <strong>of</strong> human semen, 2010<br />
2 Gravance et al., 1996<br />
3 Gravance et al., 2009<br />
4 Rijsselaere et al., 2004<br />
5 García-Herreros et al., 2007<br />
Literature on equine semen reports a wide variety both in CASA motility settings (Loomis<br />
<strong>and</strong> Graham, 2008; Ortega-Ferrusola et al., 2009; Vidament et al., 2009; Waite et al.,2008) as well as<br />
in the use <strong>of</strong> different chambers in combination with CASA systems. The chambers most frequently<br />
used are a 20 µm deep Leja chamber (Hoogewijs et al., 2010; Len et al., 2010; Ortega-Ferrusola et al.,<br />
2009; Waite et al., 2008), the 20 µm deep Cell-vu chamber (Almeida <strong>and</strong> Ball, 2005; Glazar et al.,<br />
2009; Spizziri et al., 2010) <strong>and</strong> the 10 µm deep Makler chamber (Johannisson et al., 2009; Kavak et<br />
al., 2003). These chambers are frequently loaded with a different volume, <strong>and</strong> sometimes only the