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Updated: Jun 4, 2026
Fluorimetric Techniques for the Assessment of Sperm Membranes
Published on: November 28, 2018
Agreement between measures of total motility and membrane integrity in stallion sperm
M L Foster1, D D Varner, K Hinrichs
1Department of Large Animal Clinical Sciences, Texas A&M University, College Station, TX, USA.
Abstract:
Increasing seminal plasma concentrations in extended stallion semen were utilized to model decreasing sperm motility over time. Level of agreement was determined between flow cytometric measurement of sperm membrane integrity, using a combination of SYBR-14 and propidium iodide, and computer-assisted analysis of sperm motility. Values for total sperm motility (TMOT;%) and membrane integrity (SMI;%) were similar (∼80%) at Time 0 within all sperm treatments. However, TMOT was lower than SMI after 24 and 48 h of storage in treatments with >20% seminal plasma. At Time 0, agreement (bias and absolute difference) between TMOT and SMI was high (-0.7 and 5.6%, respectively), but decreased after 24 (10.8 and 15.1%, respectively) and 48 h (23.0 and 23.8%, respectively) of cooled storage as motility declined more rapidly than SMI. We concluded that TMOT and SMI measured separate aspects of sperm quality.
Insights
Seminal plasma affects stallion sperm quality. Total sperm motility (TMOT) and sperm membrane integrity (SMI) measurements diverge over time, indicating they assess different aspects of sperm health during storage.
Area of Science:
- Reproductive Biology
- Animal Science
- Veterinary Medicine
Background:
- Extended stallion semen requires careful quality assessment for artificial insemination.
- Seminal plasma composition can influence sperm viability during cryopreservation and storage.
- Accurate methods are needed to evaluate sperm motility and membrane integrity over time.
Purpose of the Study:
- To model the decline of sperm motility in extended stallion semen.
- To compare flow cytometric sperm membrane integrity (SMI) with computer-assisted sperm motility (TMOT) analysis.
- To determine the agreement between TMOT and SMI during cooled storage.
Main Methods:
- Utilized increasing seminal plasma concentrations to model decreasing sperm motility.
- Employed flow cytometry with SYBR-14 and propidium iodide for SMI measurement.
- Used computer-assisted analysis for total sperm motility (TMOT) assessment.
- Compared TMOT and SMI values at 0, 24, and 48 hours of cooled storage.
Main Results:
- Total sperm motility (TMOT) and sperm membrane integrity (SMI) were similar (~80%) at Time 0.
- TMOT declined more rapidly than SMI in treatments with >20% seminal plasma after 24 and 48 hours.
- Agreement between TMOT and SMI decreased significantly over the 48-hour storage period.
Conclusions:
- Total sperm motility and sperm membrane integrity measure distinct parameters of stallion sperm quality.
- Seminal plasma concentration impacts the divergence between TMOT and SMI during storage.
- Flow cytometry for SMI offers a complementary assessment to motility analysis in extended stallion semen.
