Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Electron microscopic study of human sperm membrane isolation

P F Levay1, N Lourens, G P Loots

  • 1Department of Obstetrics and Gynaecology, University of Pretoria, South Africa.

Journal of Assisted Reproduction and Genetics
|July 1, 1994
PubMed
Summary

This study optimized human sperm membrane isolation using sonication and sucrose gradient centrifugation. The best method requires 3 x 15 sec sonication and yields pure sperm membranes with minimal contamination.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Ultrastructure of the cardiac and pyloric glands of the gastric mucosa of the South African hedgehog, Atelerix frontalis.

Journal of morphology·2018
Same author

Expression of the H-subunit and L-subunit of ferritin in bone marrow macrophages and cells of the erythron during cellular immune activation.

Blood cells, molecules & diseases·2011
Same author

Pro- and anti-inflammatory cytokines during immune stimulation: modulation of iron status and red blood cell profile.

Mediators of inflammation·2011
Same author

Causes of high erythrocyte sedimentation rates in an inpatient population.

South African medical journal = Suid-Afrikaanse tydskrif vir geneeskunde·2005
Same author

Blood pressure and melatonin in chronic renal failure.

Clinical nephrology·2001
Same author

Two roles for caveolae in the cardiac myocyte?

South African medical journal = Suid-Afrikaanse tydskrif vir geneeskunde·1998

Area of Science:

  • Reproductive Biology
  • Cell Biology
  • Biochemistry

Background:

  • Human sperm membranes are crucial for fertilization.
  • Previous methods for isolating sperm membranes often resulted in cellular contamination.
  • A pure and homogeneous sperm membrane preparation is needed for detailed biochemical and functional studies.

Purpose of the Study:

  • To develop a simple and effective method for isolating pure, homogeneous human sperm membranes.
  • To optimize sonication parameters for sperm membrane isolation.
  • To evaluate different sucrose gradient centrifugation techniques for removing cellular contaminants.

Main Methods:

  • Spermatozoa were sonicated using ultrasound, with varying sonication times tested (3x5, 3x15, and 180 seconds).

Related Experiment Videos

  • Sonicated samples were centrifuged, and the supernatant was layered on either a sucrose cushion or a discontinuous sucrose gradient.
  • Fractions were analyzed via electron microscopy after fixation with tannic acid and glutaraldehyde.

    • Main Results:

    • Optimal sonication time for membrane isolation was determined to be 3 x 15 seconds.
    • Isolation using a simple sucrose cushion proved inadequate due to significant cellular contamination.
    • The most effective method involved isolating sperm membranes from the sucrose interface between 0.75 M and 1.05 M sucrose.

    Conclusions:

    • The developed method offers a simple approach to obtaining pure human sperm membranes.
    • A significant drawback is the large quantity of spermatozoa required for effective membrane purification.
    • This technique provides a valuable tool for further research into sperm membrane composition and function.