Comparison of different counting chambers using a computer-assisted semen analyzer

Figures & data

Figure 1. Validation of the counting chambers. Validation of the counting chambers against the standard latex bead concentrations (A) 23.5 × 10⁶/mL sample and (B) 51.7 × 10⁶/mL sample. Significantly higher counts of two kinds of standard latex bead concentrations were obtained from Makler than from other counting chambers. Similar descripitive and comparative stastical analysis of all counting chambers with CASA in fresh and separated sperm concentration test (C and D); (C) fresh sperm sample and (D) separated sperm sample. Assessments of sperm motility by various counting chambers with CASA (E and F); (E) fresh sperm sample and (F) separated sperm sample. Significantly higher motility percent of fresh sperm from dropping loaded coverslips (including Makler, Slide, and Cover) than that of capillary-loaded (including GoldCyto and Leja). The standard deviations in C, D, E, and F (including in Figure 2), are relatively higher because of the statistical data from different semen samples. CON: concentration, MOT: motility.

Table1. The standard latex bead concentrations (mean ± SD × 10⁶/mL) obtained by different counting chambers.

	Makler	GoldCyto	Slide	Cover	Leja
23.5 (×10^6/mL), n = 31	40.46 ± 3.92 (9.69%)#	21.5 ± 2.46 (11.44%)	23.41 ± 5.18 (22.13%)	26.19 ± 5.58 (21.31%)	25.23 ± 1.62 (6.42%)
51.7 (×10^6/mL), n = 30	77.11 ± 3.73 (4.84%)	48.53 ± 3.09 (6.37%)	51.30 ± 5.04 (9.82%)	49.68 ± 5.02 (10.10%)	51.66 ± 2.70 (5.23%)
	Makler-GoldCyto	Makler-Slide	Makler-Cover	Makler-Leja	GoldCyto-Slide
23.5 (×10^6/mL)	P < 0.001^&	P < 0.001	P < 0.001	P < 0.001	P = 0.493
51.7 (×10^6/mL)	P < 0.001	P < 0.001	P < 0.001	P < 0.001	P = 0.122
	GoldCyto-Cover	GoldCyto-Leja	Slide-Cover	Slide-Leja	Cover-Leja
23.5 (×10^6/mL)	P < 0.001	P < 0.001	P = 0.248	P = 0.491	P = 0.952
51.7 (×10^6/mL)	P = 0.961	P = 0.001	P = 0.901	P = 0.464	P = 1.000

LSD post-hoc test was used for multiple comparisons between groups. #: The CV% is shown in brackets; &: P valve in bold represents statistically significant.

Table 2. Descriptive and comparative sperm concentrations (CON) (×10⁶/mL), motility (MOT %) assessment, and kinematic measurements from 56 patients by different counting chambers (mean ± SD).

	Fresh sperm
Parameters	Makler	GoldCyto	Slide	Cover	Leja
CON × 10^6/mL	103.00 ± 54.18	55.04 ± 25.73	55.75 ± 27.20	53.68 ± 25.78	59.36 ± 27.46
MOT %	61.63 ± 18.55	54.83 ± 16.99	68.03 ± 18.38	63.35 ± 20.01	56.35 ± 17.31
PR %	36.24 ± 16.95	26.89 ± 13.77	37.97 ± 15.98	34.06 ± 16.82	25.16 ± 11.82
NP %	25.40 ± 6.67	27.93 ± 7.15	30.06 ± 7.79	29.28 ± 7.79	31.19 ± 8.25
IM %	8.37 ± 18.55	45.17 ± 16.99	31.91 ± 18.38	36.65 ± 20.01	43.65 ± 17.31
VCL μm/s	38.11 ± 7.63	33.93 ± 7.14	38.01 ± 7.30	35.67 ± 7.26	32.30 ± 6.17
VSL μm/s	18.28 ± 5.64	15.05 ± 4.40	16.67 ± 4.86	14.98 ± 4.30	14.15 ± 3.68
VAP μm/s	25.36 ± 5.99	21.84 ± 4.87	24.36 ± 5.24	22.67 ± 4.87	20.78 ± 4.05
LIN %	47.30 ± 7.22	43.86 ± 6.07	43.20 ± 6.96	41.69 ± 6.25	43.52 ± 5.82
STR %	71.06 ± 6.50	67.98 ± 6.29	67.34 ± 7.29	65.32 ± 6.57	67.42 ± 6.18
WOB %	66.27 ± 5.07	64.34 ± 4.69	63.86 ± 4.66	63.59 ± 4.84	64.39 ± 4.50
	Separated sperm
Parameters	Makler	GoldCyto	Slide	Cover	Leja
CON × 10^6/mL	35.00 ± 24.07	17.58 ± 12.49	20.24 ± 15.64	18.73 ± 12.41	16.32 ± 11.55
MOT %	90.02 ± 7.15	92.06 ± 7.00	81.87 ± 12.95	82.22 ± 13.04	93.22 ± 7.37
PR %	80.58 ± 11.22	83.12 ± 12.71	63.09 ± 20.05	64.74 ± 19.71	84.84 ± 12.07
NP %	8.96 ± 5.19	7.74 ± 3.80	18.83 ± 11.96	16.19 ± 9.93	7.75 ± 3.77
IM %	10.45 ± 7.52	9.15 ± 10.30	18.10 ± 12.93	19.07 ± 14.33	7.41 ± 9.09
VCL μm/s	99.03 ± 15.44	94.86 ± 14.65	72.90 ± 22.78	72.23 ± 24.03	89.92 ± 18.12
VSL μm/s	46.43 ± 10.63	39.01 ± 10.74	23.79 ± 10.42	21.66 ± 10.41	42.56 ± 9.16
VAP μm/s	58.18 ± 10.81	51.84 ± 9.98	35.31 ± 11.94	33.65 ± 12.34	53.10 ± 10.15
LIN %	46.89 ± 8.30	40.97 ± 9.40	32.32 ± 9.68	28.70 ± 10.74	47.76 ± 7.47
STR %	79.20 ± 6.03	74.11 ± 8.37	65.26 ± 11.42	60.38 ± 14.93	79.90 ± 5.57
WOB %	58.83 ± 6.88	54.67 ± 6.96	48.51 ± 6.99	45.36 ± 9.77	59.47 ± 5.99

LSD post-hoc test was used for multiple comparison between groups. p values are shown in Supplementary Table 1. PR: progressive motility; NP: non-progressive motility; IM: immotile spermatozoa; VCL: curvilinear velocity; VSL: straightline velocity; VAP: average path velocity; LIN: linearity; STR: straightness; WOB: wobble.

Figure 2. Measurements of motility and kinematics of fresh sperm under the different concentrations (million/mL). The higher the sperm concentration, the higher the percent of sperm motility and non-progressive. PR: progressive; NP: non-progressive; CON: concentration.

Figure 3. Ancillary tests of 32 fresh sperm samples using three counting chambers, including GoldCyto, Slide, and Cover. (A) Analysis of concentrations (p > 0.05) and (B) measurements of motility (p value between GoldCyto and Slide less than 0.05, p value between GoldCyto and Cover less than 0.05, p value between Slide and Cover more than 0.05). CON: concentration; MOT: motility.