精子计数板深度与精子浓度的关系研究

The effects of the depth of sperm chamber on sperm concentration detection

目的：探讨精子计数板深度与精子浓度检测结果之间的关系，以及相应的校正方法。方法利用Filmetrics间隙测量仪精确测量5种不同深度的Geoffrey精子计数池，然后按照WHO5手册的要求利用瑞祺CFT-9201型精子质量检测分析系统分析高[标称值为50×106/ml（43.9~57.0×106/ml）]、低浓度[标称值为30×106/ml （26.3~34.2×106/ml）]的微球悬浮液（质控珠）的微球浓度，以及31例精液标本的精子浓度。并分析不同精子计数池深度所得结果的相关性、相对偏差以及校正系数。结果5种精子计数池的深度分别为8.1、9.1、10.2、11.1、11.9μm。不同深度的精子计数池计数的低、高浓度质控珠浓度结果均有显著差异（P＜0.01），但低、高浓度质控珠浓度与计数池深度均呈显著正相关（r均为0.997，P均＜0.01）。以10.2μm的计数板为基准（100%），5种计数板深度的相应校正系数分别为：1.26（10.2/8.1）、1.12（10.2/9.1）、1.00（10.2/10.2）、0.92（10.2/11.1）和0.86（10.2/11.9），校正后的各计数池相对偏差均低于5%。31例精液标本用不同深度的精子计数池进行检测，精子浓度结果类似于质控珠悬液。结论在精子浓度分析中，精子计数池的深度非常关键，使用不合格的计数板，将导致检测结果产生误差，这种误差和计数板深度误差成正比。因此，精子计数板深度应该进行定期校验，并应以相应的校正系数对检测结果进行纠正。

Objective To investigate the effects of the depth of sperm chamber on sperm concentration detection, and explore the corresponding correction method. Methods The depth of 5 sperm chambers were determined by the Filmetrics F20 Spectral Reflectance Thin-Film Measurement System. Then, the bead concentrations for low （nominal value was 30× 106/ml [26.3~34.2×106/ml]） and high （nominal value was 50×106/ml [43.9~57.0×106/ml]） concentration of bead solutions and sperm concentrations from 31 semen samples were detected by Ruiqi CFT-9201 computer-aided sperm analysis （CASA） system according to the standard of WHO5 manual, and the correlation, relative deviation and the coefficient of correction for the results obtained from 5 sperm chambers with different depth were analyzed. Results The depths of 5 sperm chambers were 8.1, 9.1, 10.2, 11.1 and 11.9 μm, respectively. There was significant difference of bead concentration between different depths of sperm chambers for low and high concentration of bead solutions （P〈0.01）. However, there was significantly positive correlation between the depth of sperm chamber and bead concentration （r=0.997,P〈0.01）. If the 10.2μm of sperm chamber was as the standard （100%）, the coefficients of correction for 8.1, 9.1, 10.2, 11.1 and 11.9 μm of sperm chambers were 1.26 （10.2/8.1）, 1.12 （10.2/9.1）, 1.00 （10.2/10.2）, 0.92 （10.2/11.1） and 0.86 （10.2/11.9）, respectively. Moreover, the relative deviations for all 5 sperm chambers were less than 5% after correction. The results from 31 semen samples were similar to that of bead solutions. Conclusion In the analysis of sperm concentration, the depth of sperm chamber is critical. The use of unqualified sperm chamber will lead to the incorrect result of sperm concentration, and the produced error is proportional to the error of sperm chamber depth. Therefore, the depth of sperm chamber should be checked regularly, and the determination result should be corrected with the corresponding coefficient of correction.