通过荧光原位杂交改进植入前基因诊断的固定方法。
Improving the fixation method for preimplantation genetic diagnosis by fluorescent in situ hybridization.
作者信息
Xu K, Huang T, Liu T, Shi Z, Rosenwaks Z
机构信息
Center for Reproductive Medicine and Infertility, Cornell University Medical College, New York, New York 10021, USA.
出版信息
J Assist Reprod Genet. 1998 Oct;15(9):570-4. doi: 10.1023/a:1022542406056.
PURPOSE
Our purpose was to modify a fixation method using Tween-20 and HCl (TH) and to compare it with a protocol using methanol and acetic acid (MA) for the improvement of preimplantation genetic diagnosis by fluorescence in situ hybridization (FISH).
METHODS
Single blastomeres were allocated to either the TH or the MA procedure. The two methods were compared to evaluate efficiency of fixation and the intensities of FISH signals.
RESULTS
With the TH method, 123 (93.9%) of 131 blastomeres were fixed, while only 95 (78.5%) of 121 were fixed with MA. Average scores for the intensity of FISH signals were significantly stronger for TH than for MA (P < 0.05). There was also a significant difference in signal intensity scores between the two methods for type-3 nuclei.
CONCLUSIONS
Our results indicate that not only are fewer nuclei lost during fixation but also stronger FISH signals can be obtained with the TH method. Thus, modified TH can improve the overall efficiency of preimplantation genetic diagnosis.
目的
我们的目的是改进一种使用吐温 - 20和盐酸（TH）的固定方法，并将其与使用甲醇和乙酸（MA）的方案进行比较，以通过荧光原位杂交（FISH）改善植入前遗传学诊断。
方法
将单个卵裂球分配到TH或MA程序中。比较这两种方法以评估固定效率和FISH信号强度。
结果
使用TH方法，131个卵裂球中有123个（93.9%）被固定，而使用MA方法时，121个中只有95个（78.5%）被固定。TH方法的FISH信号强度平均得分显著高于MA（P < 0.05）。两种方法在3型细胞核的信号强度得分上也存在显著差异。
结论
我们的结果表明，TH方法不仅在固定过程中丢失的细胞核较少，而且能获得更强的FISH信号。因此，改进后的TH方法可以提高植入前遗传学诊断的整体效率。
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