Lacquet F A, Kruger T F, Du Toit T C, Lombard C J, Sanchez Sarmiento C A, De Villiers A, Coetzee K
Clinical Laboratory, St. Blasius Hospital, Dendermonde, Belgium.
Arch Androl. 1996 Mar-Apr;36(2):133-8. doi: 10.3109/01485019608987089.
The purpose of this study was to standardize slide preparation and staining procedures to improve the efficiency and effectivity of the IVOS system on normal sperm morphology readings with regard to the strict criteria. Semen samples from patients attending the Reproductive Biology Unit, Tygerberg Hospital, were used. In experiment 1, five different Diff-Quik staining procedures, including the standard procedure, were evaluated on each of 22 patients and the effect of slide preparation within 1 h or more than 5 h after collection and the effect of immediate fixation versus fixation after 24 h were observed. In experiment 2, the manual evaluation time per slide (n = 20) by two technicians was compared with the time taken by computer. In experiment 1 the median % normal for the 5 different staining procedures was 6, 6.5, 9.5, 8.5, and 5.5%. No significant difference was found between the different staining procedures (p = .60, nonparametric Friedman test). In experiment 2 the mean time for manual assessment by two technicians was 3 min:6 s and 3 min:53 s per slide as compared to 4 min:39 s by computer. For experiment 1, slides can be prepared immediately or after 5 h. Fixation time also does not interfere with the computer's ability to identify normal forms. For experiment 2, the IVOS system is competitive regarding assessment time. Standardization of optimum staining procedures is important to ensure repeatability and comparability. Therefore, slides should be prepared immediately after liquefaction and fixed immediately after air drying.
本研究的目的是规范载玻片制备和染色程序，以提高IVOS系统在严格标准下对正常精子形态读数的效率和有效性。使用了来自泰格堡医院生殖生物学科患者的精液样本。在实验1中，对22名患者中的每一位评估了包括标准程序在内的五种不同的Diff-Quik染色程序，并观察了采集后1小时内或5小时以上制备载玻片的效果以及立即固定与24小时后固定的效果。在实验2中，比较了两名技术人员对每张载玻片（n = 20）的手动评估时间与计算机所用时间。在实验1中，五种不同染色程序的正常百分比中位数分别为6%、6.5%、9.5%、8.5%和5.5%。不同染色程序之间未发现显著差异（p = 0.60，非参数弗里德曼检验）。在实验2中，两名技术人员的手动评估平均时间为每张载玻片3分06秒和3分53秒，而计算机为4分39秒。对于实验1，载玻片可以立即制备或在5小时后制备。固定时间也不会干扰计算机识别正常形态的能力。对于实验二，IVOS系统在评估时间方面具有竞争力。优化染色程序的标准化对于确保可重复性和可比性很重要。因此，精液液化后应立即制备载玻片，空气干燥后应立即固定。
