CAJ | 학술논문

用曙红 Y、台盼蓝染料对马氏珠母贝(Pinctada martensii)精子进行染色，通过对不同制片方法、不同的染色时间和不同染料浓度的染色效果进行比较，筛选出可通过染色特征来检测精子死活及活精率的适宜方法，结果显示：(1)经染色后，死、活精子呈现出明显不同的特征：活精子均不着色，头部呈梨形、无色透明状；死精子着红色(曙红Y染色)或蓝色(台盼蓝染色)，体积变大，头部近圆形。(2)曙红Y、台盼蓝最适染色浓度分别为0.04%和0.2%；最佳染色时间均为15min。(3)检测到的精子死亡率即实测精子死亡率，与“理论死亡率”之间有明显的相关性，相关系数为R=0.99(P＜0.01)，并且两种染色法的观察结果一致，表明这些检测结果客观可信；(4)染料对活精子毒性的检测显示，染色45min内，精子死亡率无显著性增加(P＞0.05)，说明在该时间范围内，染液对精子的毒性较小，精子存活率的检测结果稳定可靠。这些研究表明，曙红 Y、台盼蓝两种染色法准确灵敏，均适宜用于马氏珠母贝精子质量的检测与评价。
용서홍 Y、태반람염료대마씨주모패(Pinctada martensii)정자진행염색，통과대불동제편방법、불동적염색시간화불동염료농도적염색효과진행비교，사선출가통과염색특정래검측정자사활급활정솔적괄의방법，결과현시：(1)경염색후，사、활정자정현출명현불동적특정：활정자균불착색，두부정리형、무색투명상；사정자착홍색(서홍Y염색)혹람색(태반람염색)，체적변대，두부근원형。(2)서홍Y、태반람최괄염색농도분별위0.04%화0.2%；최가염색시간균위15min。(3)검측도적정자사망솔즉실측정자사망솔，여“이론사망솔”지간유명현적상관성，상관계수위R=0.99(P＜0.01)，병차량충염색법적관찰결과일치，표명저사검측결과객관가신；(4)염료대활정자독성적검측현시，염색45min내，정자사망솔무현저성증가(P＞0.05)，설명재해시간범위내，염액대정자적독성교소，정자존활솔적검측결과은정가고。저사연구표명，서홍 Y、태반람량충염색법준학령민，균괄의용우마씨주모패정자질량적검측여평개。
The survival situation and rate of sperms of pearl oyster Pinctada martensii was detected by being stained with eosin Y stain and typan blue respectively, the suitable ways were selected by comparing the effect of different sampling method, staining duration and dye concentration. The results showed that (1) there were significant differences of staining characters between dead and live spermatozoas:the live ones were transparent, spherical and became bigger, while the dead ones were stained easily by either dye; (2) the optimum staining duration of both dyes was 15min, and the optimum dye concentration was 0.04%eosin Y and 0.2%trypan blue respectively;(3) the theoretical and real mor-talities were correlated with coefficient R=0.99 (P<0.01) and the rates detected by both eosin Y stain and trypan blue showed the same result;(4) the test of dye toxicity to live sperms showed that the mortality did not increase significantly with P>0.05 with as staining duration prolonging to 45min. These results meant that dye solution was less toxic to sperms, and it was more credible to detect the survival rate of sperms within staining duration of 45min. All these results suggested that staining with either eosin Y or trypan blue was suitable for evaluation of sperm survival rate of Pinctada martensii and the results were sensitive and accurate.