Dxz1 dyz3

Six placental samples from a male fetus were used to identify infiltrating cell origin by fluorescence in situ hybridization (FISH). Slides were placed in a 90°C oven for 15 minutes. Slides were then deparaffinized with xylene (2 times, 15 minutes each) at room temperature (RT), dehydrated in 100% ethanol for 5 minutes at RT, and placed in 10mM Citric Acid (pH 6.0) and microwaved for 10 minutes. The slides were then dehydrated using an ethanol series (70, 85, 100%) 2 minutes each at RT. Working solution of DXZ1/DYZ3 (Abbott Laboratories, Des Plaines, IL, USA) was made by mixing 1 µL of concentrated DXZ1/DYZ3 probe with 9 µL of LSI/WCP hybridization buffer (Abbott Laboratories). The working solution was applied to the target areas, cover slipped, co-denatured with a ThermoBrite (Abbott Laboratories) at 83°C for 5 minutes, and hybridized overnight in a 37°C humidified oven. Following hybridization, slides were soaked in RT 2xSSC/0.1% NP-40 to remove coverslips, placed in 2xSSC/0.1% NP-40 at 74°C for 2 minutes and then placed into RT 2xSSC/0.1% NP-40 for 2 min. The slides were stained with 4’−6,-diamidino-2-phenylindole (DAPI) (Vector Laboratories) and cover slipped. Tissue samples were scanned in their entirety and the qualitative result was determined based on observed signal patterns by CytoVision (Leica Biosystems, Wetzlar, Germany).