Examination and processing of human semen - libdoc.who.int ...

228 APPENDIX 2 Equipment and safety
condoms: spermicide-free, non-toxic (optional);
dilution vials;
dissecting microscope (optional; for collecting hamster oocytes);
filter paper, 90 g/m 2 (for filtering stains);
fluorescence microscope and objectives (optional; for high-sensitivity sperm
concentration measurements and acrosome reaction tests);
haemocytometers: improved Neubauer or alternative, 100 m deep, with thick
coverslip (thickness number 4, 0.44 mm);
incubator (37 °C), preferably with 5% (v/v) CO 2 (optional);
laboratory film: self-sealing, mouldable;
laboratory multi-key counter (six or nine keys);
large-volume counting chamber (optional; for assessing low sperm
concentrations);
luminometer (optional; for ROS assay);
microscope slides:
– with ground glass or textured writing surface and coverslips (thickness
number 1.5, 0.16–0.19 mm);
– plain slides for pulling a drop of semen over another slide to make semen
smears;
pen/pencils:
– for writing on frosted glass slides; a pencil with lead of softness HB
(American rating number 2) is adequate;
– a wax/grease pencil (delimiting pen—optional; for limiting the area of antibody
solution on a slide);
– permanent marker pen;
pH (ISFET) electrode (optional; for viscous semen samples);
pH paper (range 6–10);
phase-contrast microscope (for estimation of sperm concentration, motility,
morphology) with at least a 50-watt light source and the following accessories
(see Appendix 3):
– ×10, ×20 (or ×25), ×40 (or ×63) positive-phase objectives, ×100 oil-immersion
objective;
– ×40 negative-phase objective (optional; for eosin vitality test);
– wide-field ×10 (or ×12.5) eyepiece (ocular);
– eyepiece reticle (for judging area of field scanned for motility);
– stage micrometer (for sperm morphology measurement);