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Improving the Fixation Method for Preimplantation Genetic Diagnosis by Fluorescent in Situ Hybridization

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Abstract

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.

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REFERENCES

  1. Handyside AH: Genetic defects in the human preimplantation embryo and the diagnosis of inherited disease. In The Biological Basis of Early Human Reproductive Failure, J van Blerkom (ed). New York, Oxford University Press, 1994, pp 345-374

    Google Scholar 

  2. Verlinsky Y, Munne S, Simpson JL, Kuliev A, Ao A, Ray P, Sermon K, Martin R, Strom C, Van Stairteghem A, Veiga A, Drury K, Williams S, Ginsberg N, Wilton L: Current status of preimplantation diagnosis. J Assist Reprod Genet 1997;14:72-75

    Google Scholar 

  3. Griffin DK, Handyside AH, Penketh RJA, Winston RM, Delhanty JD: Fluorescent in-situ hybridization to interphase nuclei of human preimplantation embryos with X and Y chromosome specific probes. Hum Reprod 1991;6:101-105

    Google Scholar 

  4. Griffin DK, Wilton LJ, Handyside AH, Winston RM, Delhanty JD: Dual fluorescent in situ hybridization for simultaneous detection of X and Y chromosome-specific probes for the sexing of human preimplantation embryonic nuclei. Hum Genet 1992;89:18-22

    Google Scholar 

  5. Griffin DK, Handyside AH, Happer JC, Wilton LJ, Atkinson G, Soussis I, Wells D, Kontogianni E, Tarin J, Geber S, Ao A, Winston RML, Delhandty JDA: Clinical experience with preimplantation diagnosis of sex by dual fluorescence in situ hybridization. J Assist Reprod Genet 1994;11:132-143

    Google Scholar 

  6. Schrurs B, Winston RML, Handyside AH: Preimplantation diagnosis of aneuploidy by fluorescent in situ hybridization: Evaluation using a chromosome 18 specific probe. Hum Reprod 1993;8:296-301

    Google Scholar 

  7. Munne S, Sultan KM, Weier H-UG, Grifo J, Cohen J, Rosenwaks Z: Assessment of numeric abnormalities of X, Y, 18, and 16 chromosomes in preimplantation human embryos before transfer. Am J Obstet Gynecol 1995;172:1191-1201

    Google Scholar 

  8. Verlinsky Y, Cieslak J, Freidine M, Ivakhnenko V, Wolf G. Kovalinskaya L, White M, Lifchez A, Kaplan B, Moise J, Valle J, Ginsberg N, Strom C, Kuliev A: Pregnancies following preconception diagnosis of common aneuploidy by fluorescent in situ hybridization. Hum Reprod 1995;10:1923-1927

    Google Scholar 

  9. Pinkel D, Landegant J, Collins C, Fuscoe J, Segraves R, Lucas J, Gray J: Fluorescence in situ hybridization with human chromosome-specific libraries: Detection of trisomy 21 and translocations of chromosome 4. Proc Natl Acad Sci USA 1988;85:9138-9142

    Google Scholar 

  10. Munne S, Dailey T, Finkestein M, Weier H-UG: Reduction in signal overlap results in increased FISH efficiency: Implications for preimplantation genetic diagnosis. J Assist Reprod Genet 1996;13:149-156

    Google Scholar 

  11. Tarkowsky AK: An air drying method for chromosome preparation from mouse eggs. Cytogenetics 1964;5:394-400

    Google Scholar 

  12. Munne S, Weier H-UG, Stein J, Grifo J, Cohen J: A fast and efficient method for simultaneous X and Y in situ hybridization of human blastomeres. J Assist Reprod Genet 1993;10:82-89

    Google Scholar 

  13. Coonen E, Dumoulin JCM, Ramaekers FCS, Hopman AHN: Optimal preparation of preimplantation embryo interphase nuclei for analysis by fluorescence in-situ hybridization. Hum Reprod 1994;9:533-537

    Google Scholar 

  14. Harper JC, Coonen E, Ramaekers FCS, Delhandty DA, Handyside AH, Winston RML, Hopman AHN: Identification of the sex of human preimplantation embryos in two hours using an improved spreading method and fluorescent in-situ hybridization (FISH) using directly labeled probes. Hum Reprod 1994;9:721-724

    Google Scholar 

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Xu, K., Huang, T., Liu, T. et al. Improving the Fixation Method for Preimplantation Genetic Diagnosis by Fluorescent in Situ Hybridization. J Assist Reprod Genet 15, 570–574 (1998). https://doi.org/10.1023/A:1022542406056

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  • DOI: https://doi.org/10.1023/A:1022542406056

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