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Related Experiment Videos

High pressure flow cytometric sorting damages sperm.

T K Suh1, J L Schenk, G E Seidel

  • 1Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins, CO 80523, USA. tksuh@colostate.edu

Theriogenology
|August 30, 2005
PubMed
Summary
This summary is machine-generated.

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Lowering the operating pressure of the MoFlo SX flow cytometer for sperm sorting to 40 pounds per square inch (psi) significantly improved sperm quality. This optimization enhances sperm motility and viability without compromising sorting efficiency.

Area of Science:

  • Reproductive Biology
  • Biotechnology
  • Animal Science

Background:

  • High-speed flow cytometry is utilized for sperm sorting, a process involving high operating pressures.
  • The standard operating pressure for the MoFlo SX flow cytometer is 50 pounds per square inch (psi) with a 70-micrometer nozzle tip.
  • Concerns exist regarding potential sperm damage due to high pressure during the sorting process.

Purpose of the Study:

  • To investigate the hypothesis that reducing sorting pressure could minimize sperm damage.
  • To determine the optimal operating pressure for sperm sorting using the MoFlo SX flow cytometer.
  • To evaluate the impact of reduced pressure on sperm quality and sorting performance in bovine and stallion spermatozoa.

Main Methods:

  • Experiments were conducted using semen from multiple bulls and stallions, processed with three MoFlo SX sorters.

Related Experiment Videos

  • Spermatozoa were sorted at various pressures, including 50 psi, 40 psi, 30 psi, and lower.
  • Sperm motility and viability were assessed post-thaw using standard assays, including SYBR-14/propidium iodide staining for viability.

    • Main Results:

    • Sorting at 30 psi significantly increased post-thaw sperm motility compared to 50 psi in bulls (48.0% vs. 40.5% at 2h).
    • Progressively motile sperm increased with decreasing pressure, reaching 39.8% at 25 psi.
    • Lowering pressure to 40 psi improved mean sperm motility (48.6%) and viability (55.7%) in bulls, and enhanced motility (40.6%) and viability (76.7%) in stallions compared to 50 psi.
    • Reducing pressure to 40 psi did not negatively impact sort rate or purity in bovine spermatozoa.

    Conclusions:

    • Lowering the operating pressure of the MoFlo SX flow cytometer from 50 psi to 40 psi demonstrably improves sperm quality.
    • Optimal sorting pressure can enhance sperm motility and viability post-sort.
    • Reduced pressure sorting offers a viable method to improve sperm quality without sacrificing sorter performance.