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Theriogenology2005; 64(5); 1035-1048; doi: 10.1016/j.theriogenology.2005.02.002

High pressure flow cytometric sorting damages sperm.

Abstract: Sexing sperm by high-speed flow cytometry subjects them to high pressure. The routine operating pressure of the MoFlo SX flow cytometer for sperm sorting for commercial production has been 50 pounds/square inch (psi), with a standard 70 microm standard nozzle tip. It was hypothesized that lowering the sorting pressure could reduce sperm damage. Therefore, a series of experiments using semen from six bulls, sorted with three MoFlo SX sorters, was conducted to determine optimal pressure. An additional experiment was done with stallion spermatozoa. In Experiment 1, sorting at 30 psi compared to 50 psi with the 70 microm nozzle tip increased sperm motility post-thaw at 30 min and 2h from 40.5 to 48.0% and 30.0 to 40.2%, respectively (P<0.05). In Experiment 2, 49, 43, 37, 31, and 25 psi resulted in 24.2, 32.8, 35.6, 37.5, and 39.8% progressively motile spermatozoa post-thaw (P<0.05). In Experiment 3, 3 pressures (50, 40, 30 psi)x2 sorting methods were further evaluated. At 50, 40, and 30 psi, respective mean sperm motilities at 30 min were 44.8, 48.6, and 49.6% (P<0.05), and percentage of live spermatozoa were 51.7, 55.7, and 57.8% (P<0.05). The improvement of post-sort sperm quality with lowered pressure was also evident in stallion spermatozoa. After sorting at 30, 40 and 50 psi were 40.6, 34.5 and 30.1% motile spermatozoa (P<0.1), and were 76.7, 72.5 and 67.8% (P0.05), and higher sperm concentration during staining increased sort rate (P<0.05). In conclusion, lowering pressure of the MoFlo SX flow cytometer for sperm sorting from 50 psi (standard pressure) to 40 psi clearly improved sperm quality without a significant decrease in sorter performance.
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Publication Date: 2005-03-02 PubMed ID: 16125550DOI: 10.1016/j.theriogenology.2005.02.002Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This paper investigates the impact of high-pressure conditions during sperm sorting using the MoFlo SX flow cytometer on the quality of sperm. The research found that reducing the sorting pressure from the standard measure of 50 psi improves sperm quality without affecting the cytometer’s performance.

Article Explanation

This research performed a series of experiments to ascertain whether reducing the pressures used in a MoFlo SX flow cytometer for sperm sorting could result in minimized sperm damage. This is critical because the cytometer’s standard operating pressure was 50 psi, which could result in a substantial degradation of sperm quality.

  • In the first experiment, lowering the pressure to 30 psi from 50 psi with a 70 micrometre nozzle tip led to a considerable increase in sperm motility post-thaw at both 30 minutes and 2 hours, demonstrating that pressure reduction is beneficial for sperm quality.

Further Findings

The study did not stop at the first experiment and conducted further tests to solidify its findings.

  • The second experiment showed that further reducing the pressure (to 49, 43, 37, 31, and 25 psi) led to a progressive increase in the percentage of motile sperm post-thaw, affirming the notion that lower pressure yields superior sperm quality.
  • The third experiment analyzed three different pressure levels combined with two sorting methods. At each respective pressure level (50, 40, and 30 psi), the mean sperm motilities increased, as did the percentage of live spermatozoa; therefore, further pressurizing the evidence of lower pressure resulting in increased sperm quality.
  • The same observations were made when the researchers carried out the same experiment in stallion spermatozoa. Lower pressure seemed to yield superior sperm quality in these animals.

Impact on Cytometer Performance

Aside from assessing sperm quality, the study also took into account the cytometer’s performance at different pressure levels.

  • In the fourth experiment, two pressures (40 and 50 psi) and two staining concentrations of bovine spermatozoa were evaluated. Notably, the research found that lowering the pressure to 40 psi did not affect the sort rate and purity of the sperm. Furthermore, increasing the sperm concentration during staining led to a higher sort rate.
  • It was concluded that reducing the MoFlo SX flow cytometer’s sperm sorting pressure from 50 psi to 40 psi improves sperm quality without significantly affecting the cytometer’s performance.

Cite This Article

APA
Suh TK, Schenk JL, Seidel GE. (2005). High pressure flow cytometric sorting damages sperm. Theriogenology, 64(5), 1035-1048. https://doi.org/10.1016/j.theriogenology.2005.02.002

Publication

Theriogenology
ISSN: 0093-691X
NlmUniqueID: 0421510
Country: United States
Language: English
Volume: 64
Issue: 5
Pages: 1035-1048

Researcher Affiliations

Suh, T K
  • Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins, CO 80523, USA. tksuh@colostate.edu
Schenk, J L
    Seidel, G E

      MeSH Terms

      • Animals
      • Cattle
      • Cell Separation
      • Cell Survival
      • Flow Cytometry / veterinary
      • Horses
      • Male
      • Pressure / adverse effects
      • Sex Determination Analysis / veterinary
      • Sperm Count
      • Sperm Motility
      • Spermatozoa / physiology

      Citations

      This article has been cited 19 times.
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