Harasymowycz J, Ball L, Seidel G E
Am J Vet Res. 1976 Sep;37(9):1053-7.
Two experiments were conducted to evaluate effects of 3 stains and 2 fixatives on morphologic features of bovine spermatozoa. In experiment 1, the morphologic features of acrosomes of raw and incubated, extended spermatozoa were evaluated after staining with Hancock's Blom's or Wells-Awa's stains or after fixation with buffered glutaraldehyde. Evaluations were done of stained smears by bright field microscopy and of fixed, unstained preparations, by differential interference contrast microscopy, using wet mounts. Raw semen samples from 1st ejaculates of 80 bulls were evaluated. The percentage of spermatozoa with intact acrosomes averaged 83.5% in unstained preparations fixed in glutaraldehyde, compared with averages of 68.1, 74.5, and 67.4% for smears stained with Hancock's, Blom's, or Wells-Awa's procedures (P less than 0.01). From these results, it appeared that procedures for preparing stained smears were detrimental to acrosomes. Although counts for other acrosomal abnormalities differed (P less than 0.01) in each treatment, patterns were inconsistent. With incubated, extended spermatozoa from 57 bulls, glutaraldehyde-fixed, unstained samples had more (55%) intact acrosomes (P less than 0.01) than did samples stained with Hancock's or Blom's procedures (24.0 and 34.7%, respectively, but the former were not significantly different from Wells-Awa-stained smears (49.3% intact acrosomes). In experiment 2, several morphologic characteristics of spermatozoa from 15 1st ejaculates of 7 bulls were evaluated after staining with Hancock's or Blom's stains or after fixation in buffered glutaraldehyde or buffered formal saline fixatives. Higher counts (P less than 0.01) of head abnormalities were found in wet, unstained fixed preparations (4.83, 4.47, 7.87, and 7.93% respectively, for Hancock's, Blom's, glutaraldehyde, and formol saline methods). There were more (P less than 0.05) separated heads on stained, dry smears (1.43, 1.23, 0.47, and 0.47%, respectively, for Hancock's, Blom's, glutaraldehyde, and formol saline procedures). Fixation with buffered glutaraldehyde resulted in higher counts (P less than 0.01) of proximal protoplasmic droplets (2.47, 1.03, 0.67, and 1.43%, respectively, for glutaraldehyde, Hancock's, Blom's, and formol saline procedures). Although not significant, the same trend was observed for distal protoplasmic droplets...
进行了两项实验，以评估3种染色剂和2种固定剂对牛精子形态特征的影响。在实验1中，用汉考克氏、布洛姆氏或韦尔斯 - 阿瓦氏染色剂染色或用缓冲戊二醛固定后，对新鲜的、孵育后的以及稀释后的精子顶体的形态特征进行评估。通过明场显微镜对染色涂片进行评估，通过微分干涉对比显微镜对固定的、未染色的制剂（使用湿片）进行评估。对80头公牛首次射精的精液样本进行了评估。戊二醛固定的未染色制剂中，顶体完整的精子百分比平均为83.5%，相比之下，用汉考克氏、布洛姆氏或韦尔斯 - 阿瓦氏方法染色的涂片平均值分别为68.1%、74.5%和67.4%（P<0.01）。从这些结果来看，制备染色涂片的程序对顶体有损害。尽管每种处理中其他顶体异常的计数不同（P<0.01），但模式并不一致。对于57头公牛孵育后的稀释精子，戊二醛固定的未染色样本的顶体完整率更高（55%）（P<0.01），高于用汉考克氏或布洛姆氏方法染色的样本（分别为24.0%和34.7%），但前者与韦尔斯 - 阿瓦氏染色涂片（顶体完整率49.3%）无显著差异。在实验2中，对7头公牛15次首次射精的精子进行了几种形态特征评估，这些精子用汉考克氏或布洛姆氏染色剂染色，或用缓冲戊二醛或缓冲甲醛生理盐水固定剂固定。在湿的、未染色的固定制剂中发现头部异常计数更高（P<0.01）（汉考克氏、布洛姆氏、戊二醛和甲醛生理盐水方法分别为4.83%、4.47%、7.87%和7.93%）。在染色的干涂片上，分离头部更多（P<0.05）（汉考克氏、布洛姆氏、戊二醛和甲醛生理盐水程序分别为1.43%、1.23%、0.47%和0.47%）。用缓冲戊二醛固定导致近端原生质滴计数更高（P<0.01）（戊二醛、汉考克氏、布洛姆氏和甲醛生理盐水程序分别为2.47%、1.03%、0.67%和1.43%）。尽管不显著，但远端原生质滴也观察到相同趋势……
