Evaluation of stallion sperm motility with ImageJ using a cell phone camera and a light microscope

ABSTRACT This study aimed to determine the accuracy of assessing stallion sperm motility using a light microscope, a cell phone camera, and a free computer-assisted semen analysis (FCASA) package for ImageJ. The total motility of frozen (n=22) and cooled (n=48) equine semen was determined by FCASA and compared to the results of subjective visual analysis (SVA) by two technicians. Frozen samples were also evaluated by a commercial computer-assisted semen analysis (CCASA) system. The Friedman test revealed no significant differences (P>0.05) between cooled samples analyzed by FCASA (38.0) and SVA (technician 1, 40.0; technician 2, 40.0), nor between frozen samples analyzed by FCASA (23.36 ± 15.9), SVA (25.5 ± 18.8 and 25.8 ± 18.5), and CCASA (25.2 ± 18.3). However, mean FCASA results were underestimated by 7.2% compared with CCASA. The correlation between FCASA and CCASA was significant and strong (P<0.0001, r=0.95). Chi-squared tests indicated that FCASA provided similar results (P=0.14) to the reference method (CCASA), but SVA had lower accuracy (P=0.04). ImageJ analysis of cell phone videos captured under a light microscope can be used for estimation of stallion sperm motility with comparable accuracy to commercial systems.

摘要 本研究旨在评估利用光学显微镜、手机摄像头以及适用于ImageJ的免费计算机辅助精液分析（free computer-assisted semen analysis, FCASA）工具包来评估种公马精子活力的准确性。研究通过FCASA检测了冷冻（n=22）与冷藏（n=48）马精液的总活力，并将结果与两名技术人员的主观视觉分析（subjective visual analysis, SVA）结果进行对比。此外，还通过商用计算机辅助精液分析（commercial computer-assisted semen analysis, CCASA）系统对冷冻样本进行了评估。弗里德曼检验结果显示：FCASA检测的冷藏样本活力（38.0）与SVA检测结果（技术人员1：40.0；技术人员2：40.0）无显著差异（P>0.05）；冷冻样本的FCASA检测结果（23.36±15.9）、SVA检测结果（25.5±18.8和25.8±18.5）与CCASA检测结果（25.2±18.3）之间同样无显著差异。不过，相较于CCASA，FCASA的平均检测结果被低估了7.2%。FCASA与CCASA的相关性显著且极强（P<0.0001，r=0.95）。卡方检验表明，FCASA与参考方法（CCASA）的检测结果无显著差异（P=0.14），但SVA的检测准确性更低（P=0.04）。基于光学显微镜下拍摄的手机视频，通过ImageJ进行分析，可用于评估种公马精子活力，其检测准确性可与商用系统相媲美。