O-021. Laser-assisted opening of the zona pellucida facilitates polar body biopsy

Montag, Markus; Ven, Katrin van der; Ven, H. van der; Delacrétaz, G.; Rink, Klaus

doi:10.1093/humrep/12.suppl_2.10

Cited by 16 publications

(22 citation statements)

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“…To biopsy 1PBs, oocytes were placed in 50 ml buffered medium drops (Gamete, Vitrolife, Göteborg) covered by mineral oil (Ovoil, Vitrolife). A 20 mm diameter hole was made in the zona pelucida (ZP) of the oocytes using a laser beam (1.48 mm noncontact diode laser; Fertilase, Medical Technologies, Montreux, Switzerland) 31 in 69 oocytes or a mechanical method (PZD) in four cases. To remove the 1PB, hand-made and commercial micropipettes (Cook, Australia) of 25 -30 mm external diameter were used.…”

Section: European Journal Of Human Geneticsmentioning

confidence: 99%

Analysis of nine chromosome probes in first polar bodies and metaphase II oocytes for the detection of aneuploidies

et al. 2003

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We used fluorescent in situ hybridisation (FISH) to detect nine chromosomes (1,13,15,16,17,18,21,22 and X) in 89 first Polar Bodies (1PBs), from in vitro matured oocytes discarded from IVF cycles. In 54 1PBs, we also analysed the corresponding oocyte in metaphase II (MII) to confirm the results; the other 35 1PBs were analysed alone as when preimplantation genetic diagnosis using 1PB (PGD-1PB) is performed. The frequency of aneuploid oocytes found was 47.5%; if the risk of aneuploidy for 23 chromosomes is estimated, the percentage rises to 57.2%. Missing chromosomes or chromatids found in 1PBs of 1PB/MII doublets were confirmed by MII results in 74.2%, indicating that only 25.8% of them were artefactual. Abnormalities observed in 1PBs were 55.8% whole-chromosome alterations and 44.2% chromatid anomalies. We observed a balanced predivision of chromatids for all chromosomes analysed. Differences between balanced predivision in 1PB and MII were statistically significant (Po0.0001, v 2 test); the 1PB was most affected. The mean abnormal segregation frequency for each chromosome was 0.89% (range 0.52-1.70%); so, each of the 23 chromosomes of an oocyte has a risk of 0.89% to be involved in aneuploidy. No significant differences were observed regarding age, type of abnormality (chromosome or chromatid alterations) or frequency of aneuploidy. Nine of the 35 patients (25.7%) whose 1PB and MII were studied presented abnormalities (extra chromosomes) that probably originated in early oogenesis. Analysis of 1PBs to select euploid oocytes could help patients of advanced age undergoing in vitro fertilization (IVF) treatment.

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Section: European Journal Of Human Geneticsmentioning

confidence: 99%

Analysis of nine chromosome probes in first polar bodies and metaphase II oocytes for the detection of aneuploidies

et al. 2003

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“…In the last years laser zona drilling has been reported to facilitate embryo hatching [9,10]. In 1998, a diode laser was used to drill a hole into the zona pellucida of mouse oocytes and to subsequently extract PB by means of a blunt-ended pipette [11].…”

Section: Introductionmentioning

confidence: 99%

Laser Microdissection as a New Approach to Prefertilization Genetic Diagnosis

Clement-Sengewald

Buchholz²,

Schütze³

2000

Pathobiology

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The genetic status of oocytes can be determined by polar body (PB) analysis. Following PB extraction, a genetic evaluation is performed. As each PB contains the complementary genetic material of the oocyte, PB analysis reveals information about its genetic status. Genetically altered oocytes may then be excluded from in vitro fertilization. The aim of our study was to evaluate laser microdissection as a tool for PB extraction purposes. Compared to the PB extraction with a sharp-ending pipette only, we could show that laser microdissection of the zona pellucida (laser zona drilling) with a UV-A laser and subsequent extraction with a blunt-ending pipette decreases the degeneration rate of oocytes. It is shown that laser pressure catapulting of extracted PB enables their contact-free transfer into tubes, thus decreasing the risk of contamination for further analysis.

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“…They were being treated for the following indications: RIF (n ¼ 7), AMA (n ¼ 5), RIF and/or AMA (n ¼ 3), and reciprocal translocation (n ¼ 1). Polar body biopsies were performed for diagnostic purposes after patients provided informed consent as described elsewhere (13), except that for zona drilling, the Octax laser was used (MTG, Altdorf, Germany), and for aspiration, custom-made capillaries were used (CustomTip 02/240, Eppendorf, Germany) (14). Further processing of polar bodies, transfer to glass slides, and fixation were performed as described elsewhere (15).…”

Section: Patients and Polar Body Biopsiesmentioning

confidence: 99%

“…Fluorescence in situ hybridization with clones RP11-120L14, located at 13q31.1, and RP11-430M15, located at 13q33.1, to chromosome preparations of a patient with the karyotype 46,XX,add(13)(p12) was performed according to standard protocols. For polar body FISH before rapid CGH analysis, a six-probe FISH set (for chromosomes 13,16,18,21,22, X) was used as described elsewhere (16). In short, co-denaturation was performed by using a hybridization oven (Hybrite; Abbott Laboratories, Abbott Park, IL), programmed at a melting temperature of 69 C for 8 minutes, followed by hybridization at 37 C overnight.…”

Section: Fluorescence In Situ Hybridizationmentioning

confidence: 99%

“…In all CGH profiles shown, the centromeres of all chromosomes, the heterochromatin block on chromosome 1, as well as the short arms of the acrocentric chromosomes (13,21,22) were shaded to indicate regions of tandem repetitive DNA sequences, which were excluded from the CGH evaluation. (A) Chromosomal CGH profiles obtained by using the standard CGH protocol (top row) and the rapid CGH protocol (bottom row) when analyzing DNA from our test system, the ovarian adenocarcinoma cell line COLO 704, carrying gains of entire large and small chromosomes as well as a gain of a chromosome arm.…”

Section: Figurementioning

confidence: 99%

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Rapid comparative genomic hybridization protocol for prenatal diagnosis and its application to aneuploidy screening of human polar bodies

Landwehr

Montag

Ven

et al. 2008

Fertility and Sterility

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O-021. Laser-assisted opening of the zona pellucida facilitates polar body biopsy

Cited by 16 publications

References 0 publications

Analysis of nine chromosome probes in first polar bodies and metaphase II oocytes for the detection of aneuploidies

Analysis of nine chromosome probes in first polar bodies and metaphase II oocytes for the detection of aneuploidies

Laser Microdissection as a New Approach to Prefertilization Genetic Diagnosis

Rapid comparative genomic hybridization protocol for prenatal diagnosis and its application to aneuploidy screening of human polar bodies

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