Centrifugation is commonly used to wash sperm; however, most washing techniques do not put sperm in a chemically defined environment. Rather, washing by centrifugation, in effect, dilutes seminal plasma components. A 0.5-mL volume of 30% (wt/vol) Accudenz was layered beneath 5 mL of 12% (wt/vol) Accudenz in a 15-mL polypropylene centrifuge tube. Diluted semen from individual males (n = 10) was ...

The purpose of this study was to characterize highly enriched human spermatozoa membrane proteins by two-dimensional electrophoresis and computer image analysis. Sperm membrane proteins were extracted by detergent solubilization from three different preparations: 1) washed semen cells following centrifugation and three wash steps in Ham's F-10 medium (the standard sperm preparation, which is co...

BACKGROUND Intracytoplasmic sperm injection (ICSI) is the primary treatment for male infertility. However for this procedure, with the exception of visual morphological selection, there is no standardization for sperm selection. Recently, the hypo-osmotic swelling test (HOST) has been proposed to potentially select sperm with intact membranes. The aim of this study is to evaluate the ability of...

OBJECTIVE To determine the effect of semen storage and separation techniques on sperm DNA fragmentation. DESIGN Controlled clinical study. SETTING An assisted reproductive technology laboratory. PATIENT(S) Thirty normoozospermic semen samples obtained from patients undergoing infertility evaluation. INTERVENTION(S) One aliquot from each sample was immediately prepared (control) for the ...

A protocol is described for individually collecting two populations of sperm cells, Svn and Sua, from pollen of Plumbago zeylanica. Pollen grains were burst in 10 mM MOPS buffer containing 0.8 M mannitol (pH 4.6). Paired sperm cells released from pollen were separated using a microinjector. Svn and Sua were then collected individually with a microinjector, based upon known size differences. Col...

The present study was conducted to assess the fertilizing potential of frozen-thawed spermatozoa, which were cryopreserved after separation on a Percoll gradient, or washed out of seminal plasma. For this purpose, binding to the zona pellucida and other characteristics of the treated sperm cells were compared with those of cryopreserved spermatozoa from the same original sample which were not m...

FEMALE GUINEA PIGS WERE INJECTED WITH THE FOLLOWING MATERIALS: homogenates of guinea pig testis in saline or in adjuvant; suspensions of washed guinea pig sperm in saline or in adjuvant; homogenates of rabbit testis in adjuvant; guinea pig sperm and rabbit sperm in adjuvant. Control animals were not injected or were injected with adjuvant alone. At various times between 15 and 39 days after inj...

Sperm ligand on Escherichia coli that agglutinates human spermatozoa has been extracted from washed E. coli cells by incubating with 3M NaCl for 12 h at 37oC under shake conditions (150 rpm). Further the sperm ligand was purified after extensive dialysis against double distilled water by step down chromatography procedures (molecular weight sieving, anion exchange). The purification procedure y...

We have been interested in determining the minimally required elements in the sperm head that are necessary in order for the paternal genome to participate in embryogenesis. We used an ionic detergent, mixed alkyltrimethylammonium bromide (ATAB), plus dithiothreitol (DTT) to remove the acrosome and almost all of the perinuclear theca, leaving only the sperm nucleus morphologically intact. We al...