title: Assessment of the separation of X- and Y-bearing sperm on albumin gradients using double-label fluorescence in situ hybridization.
abstract: OBJECTIVE
  To determine the ratio of X- to Y-bearing human spermatozoa in fractions isolated from discontinuous albumin gradients.
  
  
  METHODS
  The proportions of X- and Y-bearing sperm were determined in neat semen samples (control) and in albumin-separated fractions from the same samples. Two albumin methods were used: a two-layer method (experiment 1) and a three-layer method (experiment 2). X- and Y-bearing sperm were identified simultaneously using chromosome-specific DNA probes and fluorescence in situ hybridization.
  
  
  METHODS
  Hospital-based university department.
  
  
  METHODS
  Healthy donors with normal semen characteristics.
  
  
  METHODS
  The proportions of haploid cells (X or Y) and cells with two sex chromosomes (XX, YY, or XY) were determined.
  
  
  RESULTS
  Labeling efficiencies were > 96% in all samples. Control samples showed a 1:1 ratio of X- to Y-bearing sperm. Fractions isolated on albumin gradients showed a slight, but statistically significant enrichment of X-bearing sperm. This was evident with both albumin methods.
  
  
  CONCLUSIONS
  Discontinuous albumin gradients do not enrich Y-bearing sperm as previously reported.
authors.affiliation: Department of Obstetrics and Gynaecology, University of Adelaide, Queen Elizabeth Hospital, Woodville, South Australia.
authors.forename: H X
authors.initials: HX
authors.lastname: Wang
authors.forename: S P
authors.initials: SP
authors.lastname: Flaherty
authors.forename: N J
authors.initials: NJ
authors.lastname: Swann
authors.forename: C D
authors.initials: CD
authors.lastname: Matthews
mesh_terms: D000782:Aneuploidy; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D015342:DNA Probes; D006238:Haploidy; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D012709:Serum Albumin; D013094:Spermatozoa / Q000648:ultrastructure*; D014960:X Chromosome; D014998:Y Chromosome
chemical_list: D015342:DNA Probes; D012709:Serum Albumin