微滴法诊断隐匿精子症结合自制金属冷冻板法保存微量精子的实验研究

Experimental study on microdroplet method combined with self-made metal freezing plate for improving sperm detection rate and recovery rate in cryptozoospermia

目的研究一种创新的寻找隐匿精子症精子的方法,结合自制金属冷冻板法冷冻保存发现的微量精子,以期为患者进一步卵细胞质内单精子注射(ICSI)提供精子保障。方法选取75例常规精液涂片未见精子的标本,每份标本充分混匀后均分为两份,分别采用传统的玻片法和创新的微滴法寻找精子,发现精子后记录精子数目、活力、圆细胞数目及检测的耗时等;微滴法发现的精子立即应用自制金属冷冻板法冷冻保存大于24 h后复苏,记录复苏后的精子数目、存活数目等。结果微滴法更容易发现精子,在精子总数小于10条、10~50条及大于50条的各级标本中,微滴法的检出例数(检出率)分别为15例(20.0%)、18例(24.0%)和10例(13.3%),均高于玻片法10例(13.3%)、12例(16.0%)和8例(10.7%),差异有统计学意义。并且微滴法结合自制金属冷冻板法可以将发现的精子冷冻保存利用,获得较高的精子复苏率(91.6%),前向运动+非前向运动(PR+NP)级精子的复苏率达到56.8%。微滴法检测平均用时3.6±0.9 min较玻片法8.5±1.2 min少。结论隐匿精子症中采用微滴法更易发现精子,结合自制金属冷冻板法冷冻发现的微量精子,有较好的复苏率,复苏后的精子可用于ICSI操作,提高精子的利用率,提升患者成为生物学父亲的机会。

Objective To evaluate the sperm detection rate and recovery rate in cryptozoospermia using microdroplet method combined with self-made metal freezing plate method for improving sperm harvesting for intracytoplasmic sperm injection(ICSI).Methods Seventy-five samples without sperm in routine semen smears were selected.Each sample was fully mixed and divided into two parts.Traditional glass slide method and microdroplet method were used to search for sperm.After the sperm was found,the sperm count,motility,number of round cells and detection time were recorded.The sperm found by the microdroplet method were immediately frozen and stored for more than 24 hours by the self-made metal freezing plate method,and the number of sperm after thawing and the number of viable sperm were recorded.Results Microdroplet method obviously improved sperm detection rate in cryptozoospermia.Among the specimens with a total number of sperm less than 10,10~50 and more than 50,the sperm detection rate of microdroplet method were 20.0%(15cases),24.0%(18 cases)and 13.3%(10 cases),respectively higher than that of slide method in 13.3%(10 cases),16.0%(12 cases)and 10.7%(8 cases),the difference was statistically significant.Moreover,the microdroplet method combined with self-made metal freezing plate method achieved a high sperm recovery rate(91.6%)and high recovery rate of PR+NP sperm(56.8%)by preserving and utilizing the obtained sperm.The average test time of microdroplet method(3.6±0.9min)was less than that of slide method(8.5±1.2 min).Conclusion Microdroplet method can improve sperm detection rate in cryptozoospermia.It combines with self-made metal freezing plate method for trace sperm frozen,and has a better recovery rate.The thawed sperm can be used for ICSI operation,which improves the utilization rate of sperm and the chances of patients becoming biological fathers.