+ Translate
+ Most Popular
Cunninghamia lanceolata plantations in China
Mammalian lairs in paleo ecological studies and palynology
Studies on technological possibilities in utilization of anhydrous milk fat for production of recombined butter-like products
Should right-sided fibroelastomas be operated upon?
Large esophageal lipoma
Apoptosis in the mammalian thymus during normal histogenesis and under various in vitro and in vivo experimental conditions
Poissons characoides nouveaux ou non signales de l'Ilha do Bananal, Bresil
Desensitizing efficacy of Colgate Sensitive Maximum Strength and Fresh Mint Sensodyne dentifrices
Administration of fluid by subcutaneous infusion: revival of a forgotten method
Tundra mosquito control - an impossible dream?
Schizophrenia for primary care providers: how to contribute to the care of a vulnerable patient population
Geochemical pattern analysis; method of describing the Southeastern limestone regional aquifer system
Incidence of low birth weights in a hospital of Mexico City
Graded management intensity of grassland systems for enhancing floristic diversity
Microbiology and biochemistry of cheese and fermented milk
The ember tetra: a new pygmy characid tetra from the Rio das Mortes, Brazil, Hyphessobrycon amandae sp. n. (Pisces, Characoidei)
Risk factors of contrast-induced nephropathy in patients after coronary artery intervention
Renovation of onsite domestic wastewater in a poorly drained soil
Observations of the propagation velocity and formation mechanism of burst fractures caused by gunshot
Systolic blood pressure in a population of infants in the first year of life: the Brompton study
Haematological studies in rats fed with metanil yellow
Studies on pasteurellosis. I. A new species of Pasteurella encountered in chronic fowl cholera
Dormancy breaking and germination of Acacia salicina Lindl. seeds
therapy of lupus nephritis. a two-year prospective study

Sample preparations are important for fluorescence in situ hybridization of cells biopsied from preimplantation embryos

Sample preparations are important for fluorescence in situ hybridization of cells biopsied from preimplantation embryos

Zygote 22(3): 300-304

ISSN/ISBN: 0967-1994

PMID: 23331574

DOI: 10.1017/s0967199412000639

Fluorescence in situ hybridization (FISH) is a cytogenetic technology used to detect chromosomal abnormalities in preimplantation human embryos. However, its efficiency is not stable due to improper sample preparation. The present study was designed to modify the current sample preparation technique and then to evaluate its efficiency in human preimplantation genetic diagnosis (PGD). Day 3 cleavage embryos as well as day 5 and 6 blastocysts were biopsied by mechanical aspiration method. In the present study, two methods were used for sample preparation of the biopsied cells. Method I was the traditional method, in which each blastomere was placed in a hypotonic solution for 5 min and then fixed on glass slides. The slides were kept at room temperature before the FISH procedures. Method II was a modified method, in which all blastomeres were placed individually in hypotonic solution drops covered by oil for at least 5 min and then fixed on slides with 0.1% Tween/HCl. After fixation, the slides were kept at -20°C for at least 30 min before the FISH procedures. The two methods were compared in terms of time consumption and proportions of blastomeres with FISH signals. In total, 329 blastomeres from day 3 embryos were fixed by Method I with an average fixation time of 8-10 min for each blastomere. By contrast, with Method II, 362 blastomeres were fixed and the average time was 3-4 min for each blastomere. After FISH, more nuclei had signals with Method II (97.2%) than with Method I (86.9%). All cells that were biopsied from blastocysts and prepared with Method II had FISH signals. However, Method I was not suitable for the fixation of multiple cells biopsied from blastocysts as cells were not traceable during the fixation. The present study indicates that proper sample preparation is critical for obtaining FISH signals in cells biopsied from preimplantation human embryos; hence these modifications can increase the efficiency of human PGD.

Please choose payment method:

(PDF emailed within 0-6 h: $19.90)

Accession: 055651432

Download citation: RISBibTeXText

Related references

Genetic diagnosis using polymerase chain reaction and fluorescent in-situ hybridization analysis of biopsied cells from both the cleavage and blastocyst stages of individual cultured human preimplantation embryos. Human Reproduction 10(1): 183-192, 1995

Muc1 in secretory phase endometrium: expression in precisely dated biopsies and flushings from normal and recurrent miscarriage patients. Human Reproduction 10(10): 2655-2662, 1995

Number of embryos biopsied as a predictive indicator for the outcome of preimplantation genetic diagnosis by fluorescence in situ hybridisation in translocation cases. Zygote 24(1): 107-114, 2016

Aneuploidy screening of preimplantation embryos using comparative genomic hybridization versus fluorescence in situ hybridization techniques. Fertility and Sterility 80(4): 873, 2003

Preimplantation aneuploidy screening using comparative genomic hybridization or fluorescence in situ hybridization of embryos from patients with recurrent implantation failure. Fertility and Sterility 80(4): 860-868, 2003

Fluorescence in situ hybridization of human polypronucleate preimplantation embryos. American Journal of Human Genetics 51(4 Suppl. ): A255, 1992

Detection of sex chromosomes by fluorescence in situ hybridization in preimplantation embryos. Journal of Assisted Reproduction and Genetics 14(8): 473, 1997

Selection of transgenic preimplantation mouse embryos using fluorescence in situ hybridization. Theriogenology 39(1): 258, 1993

Fluorescence in-situ hybridization of sex chromosomes in spermatozoa and spare preimplantation embryos of a Klinefelter 46,XY/47,XXY male. Human Reproduction 15(2): 440-444, 2000

A retrospective study of preimplantation embryos diagnosed with monosomy by fluorescence in situ hybridization (FISH). Cytogenetic and Genome Research 114(3-4): 359-366, 2006

Analysis of chromosome mosaicism in preimplantation embryos by using 2 sequential rounds of fluorescence in situ hybridization. Zhonghua Yi Xue Yi Chuan Xue Za Zhi 24(6): 706-708, 2007

Preimplantation genetic diagnosis for aneuploidy on 381 embryos from 39 initiated cycles using fluorescence in situ hybridization. Fertility and Sterility 80(Suppl 3): S197, 2003

Simultaneous detection of chromosomes X, Y, 13, 18, and 21 by fluorescence in situ hybridization in blastomeres obtained from preimplantation embryos. Journal of Assisted Reproduction and Genetics 15(5): 314-319, 1998

Numerical abnormalities for chromosomes 1, 13, 16, 18, 21, 22, X and Y from 59 preimplantation embryos using multi-color fluorescence in situ hybridization. American Journal of Human Genetics 65(4): A178, 1999

An improved technique for molecular cytogenetic analysis of human preimplantation embryos with fluorescence in situ hybridization. Journal of Reproductive Medicine 41(6): 379-383, 1996