Intra- and inter-laboratory variability in the assessment of sperm morphology by strict criteria: impact of semen preparation, staining techniques and manual versus computerized analysis

We designed prospective studies to compare manual and computerized analysis of sperm morphology by strict criteria using different semen processing an d staining techniques. A total of 54 semen samples were studied; slides wer e prepared from each subject from liquefied semen and after washing, and st ained with Diff-Quik(R) or Papanicolaou, An intra-laboratory, blind assessm ent was performed manually (two observers) and using a computerized analyse r (two readings). This demonstrated a very good correlation between manual analysis of liquefied and washed samples with both staining techniques [int raclass coefficient (ICC) = 0.93 and 0.83], Greater agreement was observed between computerized readings (washed samples) of Diff-Quik(R) (ICC = 0.93) than of Papanicolaou-stained slides (ICC = 0.66), An excellent intra-labor atory correlation was observed for within-computer readings (ICC = 0.93), T here was moderate agreement between inter-laboratory computer readings (two centres, ICC = 0.72), Although there was lower inter-laboratory agreement for manual and manual versus computer readings, overall results of all manu al and computer analyses showed good agreement (ICC = 0.73), Diff-Quik(R) s taining is reliable for both manual (liquefied) and computer (washed) analy sis of strict sperm morphology, Intra- and inter-computer analyses using th is method reached satisfactory levels of agreement. There is still high int er-laboratory variability for the manual method.