Abstract
To modify a fixation method improving the intensity and clarity of the single blastomeric signal detection by fluorescence in situ hybridization (FISH) in preimplantation genetic diagnosis. 333 cycles of assisted reproduction with preimplantation genetic diagnosis FISH (PGD-FISH) performed in our hospital were analyzed and a total of 3452 single blastomeres were obtained. For the conventional fixation method, the blastomeres were kept in 0.1% sodium citrate with 0.2 mg/ml bovine serum albumin (BSA) for 2–5 min. FISH was performed and the internal relationship between embryo quality and fixed rate, signal detection rate, and signal determination rate was explored. With the modified method, 91.54% of blastomeres were fixed, while 88.30% were fixed with the conventional method. The signal detection rate was significantly increased for the modified group than for the conventional group (compared 98.53% with 94.78%, P < 0.001). Especially, the signal determination rate also showed a significant difference between the two methods (compared 90.51% with 74.17%, P < 0.001). After the development of the fixation method, the fixation efficiency and the signal determination rate were greatly improved, providing more definite diagnosis for the patient. It will hopefully allow more assisted reproduction programs to offer their patients preimplantation genetic diagnosis with FISH.
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This study was supported by The National Key Research and Development Program of China (2017YFC1001600), National Natural Science Foundation of China (81871168) and The Fundamental Research Funds of Shandong University.
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Yu, G., Ma, S., Zhu, Y. et al. Application of improved single blastomere fixation technique in preimplantation genetic diagnosis. Cytotechnology 72, 217–226 (2020). https://doi.org/10.1007/s10616-020-00371-1
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DOI: https://doi.org/10.1007/s10616-020-00371-1