一种非侵入性、运动独立性的精子分选方法和技术，用于鉴定和取回单个有活力但非运动的精子，用于胞浆内精子注射。

A noninvasive, motility independent, sperm sorting method and technology to identify and retrieve individual viable nonmotile sperm for intracytoplasmic sperm injection.

机构信息

Department of Urology, University of California-San Francisco, San Francisco, California 94143, USA.

出版信息

J Urol. 2010 Dec;184(6):2466-72. doi: 10.1016/j.juro.2010.08.026. Epub 2010 Oct 16.

DOI:10.1016/j.juro.2010.08.026

PMID:20952004

Abstract

PURPOSE

For intracytoplasmic sperm injection in the absence of sperm motility it can be virtually impossible to distinguish viable from nonviable sperm. A reliable means to identify viable nonmotile sperm is needed and would likely improve the intracytoplasmic sperm injection success rate. Optoelectronic tweezers are a new technology that uses light induced dielectrophoresis fields to distinguish individual live cells from dead cells. We assessed the ability of optoelectronic tweezers to distinguish viable from nonviable individual nonmotile human sperm.

MATERIALS AND METHODS

Fresh semen specimens from 6 healthy men were suspended in an isotonic sucrose/dextrose solution and incubated with 0.4% trypan blue dye (Sigma-Aldrich®). Within 15 minutes we randomly selected 5 motile and 50 nonmotile sperm, including 25 trypan negative, followed by 25 trypan positive sperm, under 200× magnification for optoelectronic tweezers assay. We recorded the individual sperm response (attraction or repulsion) to the optoelectronic tweezer field and trypan staining status.

RESULTS

From each subject 55 unwashed sperm were individually assayed for a total of 330. All motile sperm were attracted to optoelectronic tweezers. Of 150 trypan negative (viable) sperm 132 (88%) were attracted to the optoelectronic tweezer field with 0.88 sensitivity (95% CI 0.82-0.93) vs that of the trypan blue assay. All 150 trypan positive (nonviable) sperm were repulsed by or neutral to the optoelectronic tweezer field with 1.0 specificity (95% CI 0.98-1.00) vs that of the trypan blue assay. Type I error equaled 0 and overall assay agreement was 94%.

CONCLUSIONS

The optoelectronic tweezer assay can distinguish viable from nonviable nonmotile viable sperm with sensitivity comparable to that of the trypan blue assay and equal specificity. Optoelectronic tweezers are a promising means of selecting sperm for intracytoplasmic sperm injection.

摘要

目的

在没有精子活动力的情况下进行胞浆内精子注射，实际上不可能区分有活力和无活力的精子。需要一种可靠的方法来识别有活力的非运动精子，这可能会提高胞浆内精子注射的成功率。光电镊子是一种使用光诱导介电泳场来区分活细胞和死细胞的新技术。我们评估了光电镊子区分有活力和无活力的个体非运动性人精子的能力。

材料和方法

将来自 6 名健康男性的新鲜精液标本悬浮在等渗蔗糖/葡萄糖溶液中，并与 0.4%台盼蓝染料（Sigma-Aldrich®）孵育。在 15 分钟内，我们在 200×放大倍数下随机选择 5 条运动和 50 条非运动精子，包括 25 条台盼蓝阴性，随后是 25 条台盼蓝阳性精子，用于光电镊子检测。我们记录了每个精子对光电镊子场的个体反应（吸引或排斥）和台盼蓝染色状态。

结果

从每个个体中未洗涤的 55 个精子进行单独检测，总共进行了 330 次检测。所有运动精子都被光电镊子吸引。在 150 条台盼蓝阴性（有活力）精子中，有 132 条（88%）被光电镊子场吸引，灵敏度为 0.88（95%置信区间 0.82-0.93），与台盼蓝检测相比。所有 150 条台盼蓝阳性（无活力）精子都被光电镊子场排斥或中性，特异性为 1.0（95%置信区间 0.98-1.00），与台盼蓝检测相比。Ⅰ类错误等于 0，总检测一致性为 94%。