BACKGROUND: In the absence of mosaicism, one of the problems of preimplantation genetic diagnosis with FISH is the occurrence of false-negative hybridization results. It has been hypothesized that missing signals are produced by spatial overlap of signals. METHODS AND RESULTS: To investigate the relation among cell density, signal overlap, and hybridization signal detection, 371 blastomeres and 4556 lymphocytes were fixed in different cellular concentrations and analyzed by FISH using probes for chromosomes X, Y, and 18, and their nuclear diameters and FISH results scored. The results showed that the lower the diameter of fixed nuclei, the higher the number of signal overlaps and missing signals. The minimum number of missing signals was obtained when lymphocyte and blastomere nuclei had 40 or more microns in diameter after fixation and FISH. Since blastomeres were fixed individually, results with blastomeres were invariably better than with lymphocytes.
BACKGROUND: In the absence of mosaicism, one of the problems of preimplantation genetic diagnosis with FISH is the occurrence of false-negative hybridization results. It has been hypothesized that missing signals are produced by spatial overlap of signals. METHODS AND RESULTS: To investigate the relation among cell density, signal overlap, and hybridization signal detection, 371 blastomeres and 4556 lymphocytes were fixed in different cellular concentrations and analyzed by FISH using probes for chromosomes X, Y, and 18, and their nuclear diameters and FISH results scored. The results showed that the lower the diameter of fixed nuclei, the higher the number of signal overlaps and missing signals. The minimum number of missing signals was obtained when lymphocyte and blastomere nuclei had 40 or more microns in diameter after fixation and FISH. Since blastomeres were fixed individually, results with blastomeres were invariably better than with lymphocytes.
Authors: J D Delhanty; D K Griffin; A H Handyside; J Harper; G H Atkinson; M H Pieters; R M Winston Journal: Hum Mol Genet Date: 1993-08 Impact factor: 6.150
Authors: Serena Emiliani; Eric Gonzalez-Merino; Marc Van den Bergh; Marc Abramowicz; Yvon Englert Journal: J Assist Reprod Genet Date: 2003-02 Impact factor: 3.412
Authors: Benjamin O'Brien; Hui Zeng; Aris A Polyzos; Kalistyn H Lemke; Jingly F Weier; Mei Wang; Horst F Zitzelsberger; Heinz-Ulrich G Weier Journal: J Histochem Cytochem Date: 2012-11-29 Impact factor: 2.479