马氏珠母贝精子质量形态评定中不同染色法染色效果的比较  

Comparison of different staining methods for morphological evaluation of the sperm quality in pearl oyster Pinctada martensii

作　　者：宾承勇[1] 王梅芳[1] 豆伟[1] 余祥勇[1] 

机构地区：[1]广东海洋大学水产学院,广东湛江524025

出　　处：《大连海洋大学学报》2012年第1期32-37,共6页Journal of Dalian Ocean University

基　　金：国家科技支撑计划项目(2007BAD29B03);广东省科技计划项目(2008B020800014);广东省重大科技兴海项目(A200708C01)

摘　　要：为筛选出适宜于马氏珠母贝Pinctada martensii精子形态分析的染色方法,采用曙红Y和结晶紫染液对不同方法制作的马氏珠母贝精液抹片进行染色,并通过显微镜观察比较染色效果。结果表明:1)采用将精液先抹片后染色(0.8 mg/mL曙红Y染液)的方法时,精子头部着色不完全,染色效果不随染色时间的延长而改善,精子形态特征不明显,故该方法不适于精子形态检测;2)将精液与20 mg/mL的曙红Y染液等体积混合后抹片,精子头部着深红色,尾部着浅红色,精子各部分结构清晰,形态特征明显,背景干净,对比明显;3)用5 mg/mL的结晶紫染液对精液进行渗透染色时,精子形态染色特征明显,头部着深紫色,尾部着浅紫色,背景干净,可较准确地判断精子的形态特征;4)依精子形态对马氏珠母贝精子进行分类,并确定了主要的畸形类型及其特点,马氏珠母贝精子畸形主要集中在精子的颈部和尾部,头部畸形情况较少;5)采用两种染色方法判定的正常精子率之间无显著差异(P<0.05),数据稳定,因此,这两种染色方法均可用于马氏珠母贝精子形态的评价中。The semen of pearl oyster Pinctada martensii was smeared and stained by two dyes Eosin Y and Crystal Violet for comparison of the staining effect and to evaluate the sperm quality in order to find out the suitable dying methods for the analysis of sperm morphology of pearl oyster.It was found that the sperm head was found to be dyed incompletely,and the sperm color was not improved with the extension of time,without obvious characteristics when the semen smear was stained with 0.8 mg/mL Eosin Y.Thus,this method is unfit for the sperm characteristic detection.After the sperm solution was mixed with 20 mg/mL Eosin Y,the sperm head was cardinal,and the tail plum,with clear parts of the sperm structure,obvious morphological characteristics and the very fair background.The obvious sperm characteristics including the dark violet sperm head and the light purple tail with the very fair background were observed in the semen stained with 5 mg/mL of Crystal Violet.So this method can be used to evaluate the sperm characteristics precisely.The main malformation was found in the pieces and tail of the sperm,few in the head according to the sperm characteristics shown by staining.The detection revealed that there were no significant differences in the evaluation of the normal sperm by the 5 mg/mL of Crystal Violet and by latter two methods（P〈0.05）,so either could be used to evaluate the sperm quality.

关 键 词：马氏珠母贝 精子 质量评价 染色 

分 类 号：S968.31[农业科学—水产养殖]