Stability of fluorochrome based assays to measure subcellular sperm functions
Stability of fluorochrome based assays to measure subcellular sperm functions作者机构：Department of Dermatology/Andrology Unit European Academy of Andrology （EAA ） Training Center University of Leipzig Leipzig 04103 Germany
出 版 物：《Asian Journal of Andrology》 (亚洲男性学杂志（英文版）)
年 卷 期：2008年第10卷第3期
页 面：455-459页
核心收录：
学科分类：1010[医学-医学技术（可授医学、理学学位）] 10[医学]
基 金：TheGermanResearchCouncil(DeutscheForschungsgemeinschaft DFGGL199/4-4)
主 题：human spermatozoa apoptosis assays caspase-3 mitochondrial membrane potential integrity CD46 semen analysis
摘 要：Aim： To evaluate the long-term stability of the fluorescence signals of new fluorescence-based semen analysis assays for clinical application. Methods： Semen samples from 87 unselected infertile patients were used to perform the following assays： （i） detection of active caspase-3 （n = 17）; （ii） integrity of the mitochondrial membrane potential （MMP） （n = 17）; （iii） externalization of phosphatidylserine （EPS） （n = 16）; and （iv） detection of intact acrosomes via CD46 （n = 37）. After the assays, 4% paraformaldehyde was added to all aliquots. The fluorescence intensity of each sample was evaluated by flow cytometry on days 0, 3, 7, 10 and 14. Results： Differences of up to ± 5% positive spermatozoa from the value measured at day 0 were estimated as acceptable deviation. The Caspase-3 FLICA^TM showed mean differences 〈 5% at day 3, 7 and 10. At day 14 the mean difference was 7.6%. In contrast, the disrupted MMP and the EPS detection showed differences 〉 5% at day 3. The CD46-FITC labeling displayed absolute differences 〈 5% CD46-positive spermatozoa at days 3, 7, 10 and 14. Conclusion： Although immediate analysis of the fluorescence signals is recommended, it is possible to evaluate caspase-3 activation up to 10 days and CD46 up to 14 days after staining of sperm. The FACS evaluation of MMP and EPS detection should be conducted on the same day.
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