The aim of this study was to evaluate the use of SYBR‐14/propidium iodide (PI) stain in a computer‐assisted spermatozoal quantification (CASQ) method of determining spermatozoal concentration in canine semen. In Experiment A, the spermatozoal concentration was measured (n = 52) with a haemocytometer and by CASQ under fluorescent illumination using green long‐pass (G‐LP) and red long‐pass filters at measurement concentrations of <25 million/ml. For the red filter, the limits of agreement between the haemocytometer and CASQ were −6.3% to 6.8% and −7.5% to 6.2% between the haemocytometer and CASQ for the G‐LP filter. For the red filter, the mean precision CVs were 2.21% ± 4.33% (mean ± 95% CI) for the haemocytometer, 2.19% ± 4.29% for CASQ and using the G‐LP filter 2.13% ± 4.18% for the haemocytometer and 2.66% ± 5.21% for CASQ. In Experiment B, spermatozoa were also examined with a green spectrum short‐pass (G‐SP) filter (n = 50) at measurement concentrations of <12.5 million/ml. The limits of agreement between the haemocytometer and CASQ were −5.4% to 7.8% using the red filter, −15.8% to 14.3% using the G‐LP filter and −13.1% to 11.3% using the G‐SP filter. The mean precision CVs for the haemocytometer and CASQ, respectively, were 2.68% ± 5.26% (mean ± 95% CI) and 1.93% ± 3.72% using the red filter and 2.01% ± 3.95% and 3.55% ± 6.95% using the G‐LP filter, and 3.96% ± 7.76% for CASQ using the G‐SP filter. Using the red filter, the agreement between the haemocytometer and CASQ and the precision of both haemocytometer methods and CASQ were better than when using green filters. The CASQ method performed using green filters produced acceptable results; however, CASQ using a red filter with PI staining alone was superior to that using green filters and SYBR‐14/PI staining.

中文翻译：

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SYBR-14和碘化丙啶染料在荧光计算机辅助的精子定量方法中的应用评估

本研究的目的是评估SYBR-14 /碘化丙啶（PI）染色剂在确定犬精液中精子浓度的计算机辅助精子定量（CASQ）方法中的使用。在实验A中，测量精子浓度（n = 52）用血细胞计数器和CASQ在荧光照明下使用绿色长通（G-LP）和红色长通滤光片在测量浓度<2500万/ ml时进行。对于红色滤光片，血细胞计数仪和CASQ之间的一致性限度是G‐LP滤光片的血细胞计数仪和CASQ之间的一致性为-6.3％至6.8％和-7.5％至6.2％。对于红色滤光片，血细胞计数仪的平均CV平均值为2.21％±4.33％（平均±95％CI），对于CASQ为2.19％±4.29％，使用G‐LP滤光片的血细胞计数仪为2.13％±4.18％和2.66对于CASQ为％±5.21％。在实验B中，还使用绿色光谱短波（G-SP）滤光片（n = 50）的测量浓度<1,250万/ ml。使用红色滤镜时，血细胞计数器与CASQ的一致性极限为–5.4％至7.8％，使用G‐LP滤镜时为−15.8％至14.3％，使用G‐SP滤镜时为−13.1％至11.3％。血细胞计数器和CASQ的平均精确CV值使用红色滤镜分别为2.68％±5.26％（平均值±95％CI）和1.93％±3.72％，使用G过滤器分别为2.01％±3.95％和3.55％±6.95％ ‐LP过滤器，使用G‐SP过滤器的CASQ为3.96％±7.76％。使用红色滤镜时，血细胞计数器与CASQ之间的一致性以及两种血细胞计数器方法和CASQ的精度均比使用绿色滤镜时更好。使用绿色滤光片执行的CASQ方法产生了可接受的结果；然而，