May 9, 2006

Influence of swim-up time on the ratio of X- and Y-bearing spermatozoa

European Journal of Obstetrics, Gynecology, and Reproductive Biology
Junhao YanYingying Qin

Abstract

The objective was to examine the separation of X- and Y-bearing spermatozoa in a modified swim-up procedure using fluorescent in situ hybridization (FISH), and to find out the influence of swim-up time on the ratio of X- and Y-bearing spermatozoa. Prospective study. Reproductive testing laboratory in a university hospital. Normal spermatozoa samples were obtained from 10 volunteers by masturbation after sexual abstinence for 3-5 days. Spermatozoa were put into 18 tubes with 0.25 ml in each, then mixed with HTF medium and centrifuged for 5 min (400 x g). The supernatant was removed and discarded and 0.5 ml HTF was added slowly along the tube wall. Motile spermatozoa were collected after swimming up in different times (from 5 up to 150 min, with a total of 17 intervals). The X- and Y-bearing spermatozoa were determined using the FISH technique. The X/Y dual-color CEP probes that were marked by fluorescein isothiocyanate (FITC) and Texas red were applied to analyze the ratio of X- and Y-bearing spermatozoa. The FISH staining slides were analyzed under an immunofluorescence microscope. About 1000-1500 spermatozoa were counted per slide. The percentages of X- and Y-bearing spermatozoa were calculated. The study results suggested tha...Continue Reading

Mentioned in this Paper

Seminal Fluid
Flow Cytometry
Immunofluorescence Assay
Fluorescent in Situ Hybridization
Ethanol
Y Chromosome
Ethanol Measurement
Imbalance
Chromosomes, Human, X
Nucleic Acid Hybridization Procedure

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