目的 以人类受精胚胎单卵裂球为对象,对传统低渗固定法进行改良,探讨一种更简便效率更高的固定方法.方法 收集30枚人类多精受精胚胎激光打孔后去除透明带,将获得的完整卵裂球按传统低渗固定法和直接固定法进行固定后行X,Y染色体双色荧光原位杂交,比较固定率和杂交率.结果 固定卵裂球共171枚.低渗固定法(A组)固定80枚,成功68枚,成功率为85.0%.直接固定法(B)组固定91枚,成功82枚,成功率为90.1%.两组差异有显著性.A组杂交率为77.5%(每卵裂球数),91.2%(每核数),B组杂交率为83.5%(每卵裂球数)92.7%(每核数).两组差异无显著性.结论 直接固定法简化了固定步骤,降低了细胞丢失率,为较理想的单个卵裂球间期核固定技术.%Objective To improve current preimplantation genetic diagnosis fixation techniques in order to find a more efficient fixation method by using polyspermy embryos. Methods At first,to remove the zona pellucida by laser. The Integrity Blastomeres were distributed into two groups: conventional hypotonic- fixationgroup and direct-fixation group. Fluorescence in situ hybridization was accomplished after fixation using chromosome X and Y probes,and then the fixation rate and the fluorescent signals were analyzed. Result 1. A total of 30 Blastomeres were analyzed and 171 Blastomeres fixated. Of the 80 Blastomeres in conventional group (A group) ,68 got fine fixation, the successful rate was 85.0%. Among 91 Blastomeres in direct-fixation group (B group) ,82 got fine fixation, the successful rate was 90. 1% . 2. In conventional group, the hybridization rate was 77. 5% (/ Blastomere) or 91. 2% (/Nuclear) . The rate were 83. 5% and 92.7% respectively with hypotonic-fixation group and the direct-fixation group. There was no Significant difference. Conclusion This improved fixation technique essentially eliminates the possibility of losing a cell during fixation and simplifies the process of fixation of blastomere for preimplatation genetic diagnosis.
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