OBJECTIVETo evaluate the accuracy and precision of 4 methods including Hemacytometer, Makler chamber, Cell-VU chamber, and computer-aided semen analysis for determining sperm concentration.

METHODSLatex bead solutions with concentrations known as (35 +/- 5) x 10(6)/ml and (18.0 +/- 2.5) x 10(6)/ml and semen samples (n = 54) were counted by the above 4 methods and the results were then compared.

RESULTSMean bead concentrations for Hemacytometer, Makler, Cell-VU chambers and CASA were (44.84 +/- 4.86), (52.36 +/- 7.78), (39.70 +/- 4.76), (28.53 +/- 2.06) x 10(6)/ml respectively for the standard solution containing (35 +/- 5) x 10(6)/ml, and (21.04 +/- 1.87), (24.54 +/- 3.67), (19.09 +/- 2.02), (14.62 +/- 0.95) x 10(6)/ml respectively for a standard solution containing (18 +/- 2.5) x 10(6)/ml. The results of Cell-VU chamber were consistently similar and close to the standard solutions, while those of Hemacytometer, Makler chambers were overestimated, and those of CASA were underestimated. The coefficients of variation for Hemacytometer, Makler, Cell-VU chambers and CASA were 10.81%, 14.86%, 12.80%, and 7.22% respectively for a higher standard solution, while 8.89%, 14.96%, 10.58%, and 6.50% respectively for a lower standard solution. CASA has the lowest CV%. When semen samples were counted, the results of Hemacytometer, Makler, Cell-VU chambers and CASA were (76.98 +/- 59.90), (63.89 +/- 53.84), (45.28 +/- 34.52), (41.96 +/- 31.93) x 10(6)/ml respectively. There wasn't any significant difference either between Cell-VU chamber and CASA (P = 0.71), or between Hemacytometer and Makler chamber (P = 0.14), while there was significant difference between Cell-VU chamber or CASA and Hemacytometer or Makler chamber (P < 0.05 or P <0.01).

CONCLUSIONWhen counting semen sample, there wasnt any significant difference between Cell-VU chamber and CASA. Each laboratory can select its own proper method for manual or computer-aided analysis.