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目的:在现有数种胚胎单细胞固定方法基础上,寻求一种稳定、理想的固定技术,并进行改良。方法:以4-细胞鼠胚(A组)和人体外受精(IVF)废弃胚胎(B组)之卵裂球为材料,分别以甲醇-冰醋酸(MA)法、吐温-盐酸(TH)改良法和MA+TH结合法进行固定,比较其各自的固定率、核形态等情况。结果:3种方法的固定率以MA法最低,且3种方法固定率差异有显著性(A组3种方法P=0.00621,B组3种方法P=0.00362);A组和B组固定后得到的核形态均与固定方法有明显相关性(P值分别为0.000和7.66×10-7);信号获得率和核信号评分与固定方法无明显关联(P值分别为0.186和0.0728)。结论:TH改良法固定后核形态不良,胞质残留率高;MA法核像直径大、质量较好,但技术要求高,难于普及;MA+TH结合法则核像较大,质量最好,技术易于掌握和推广。
OBJECTIVE: To seek a stable and ideal immobilization technique based on the existing single embryo single cell fixation methods. Methods: The blastomeres of 4-cell mouse embryos (group A) and IVF (group B) Improved method and MA + TH combination method to fix, compare their respective fixed rate, nuclear morphology and so on. Results: The fixation rates of the three methods were the lowest with MA method, and there was significant difference between the three methods (P = 0.00621 in group A, and 0.00362 in group B) The obtained nuclear morphology was significantly correlated with the fixation method (P = 0.000 and 7.66 × 10-7, respectively). The signal acquisition rate and nuclear signal score were not significantly correlated with the fixation method (P = 0.186 and 0.0728, respectively). Conclusions: The nuclear morphology of TH was poor after fixation, and the residual rate of cytoplasm was high. The diameter of MA nuclear image was large and the quality was good, but the technique was difficult to popularize. The MA + Technology is easy to grasp and promote.