Evaluierung durchflusszytometrischer Verfahren zur Beurteilung der Qualität von kryokonserviertem Hengstsperma Tierärztliche Fakultät - Digitale Hochschulschriften der LMU - Teil 01/07
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The aim of these examinations was to check various flow cytometric methods to establish whether they are suitable objective methods for assessing the quality of cryopreserved sperm from stallions. The plasma membrane integrity, the mitochondrial membrane potential, the acrosomal status and the integrity of the chromatin structure in cryopreserved sperm from stallions were assessed. For this purpose various fluorescent stainings were carried out both immediately after the semen samples were thawed and after a three-hour period of incubation at 37 C with SYBR® 14, JC-1 and SYTO® 17 / propidium iodide / FITC-PNA and a sperm chromatin structure analysis (SCSA™) and were then evaluated.
The sperm quality parameters obtained by flow cytometry showed good reproducibility. The results of measurements of three different straws of an ejaculate which were carried out independently of each other did not differ significantly. The intra-class correlation coefficients were between 0.88 and 0.98. The flow cytometric tests evaluated in this examination were therefore considered to be reliable.
In order to demonstrate the correlations between the routine examination methods for assessing the sperm quality and the analysis processes evaluated in this study, the viability and the morphology of the sperms after thawing were assessed by light microscopy using bromide phenol blue nigrosine smears. A computer controlled motility analysis was also carried out. Moderate correlations (0.60 > r > 0.58; p 0.96; p 0.05) with the fertility of the stallions. On the other hand, a negative relationship was established between the integrity of the chromatin structure and the fertility of the stallions (r = - 0.51 / - 0.59).
This study shows that the flow cytometric assessment of the quality of cryopreserved sperm from stallions represents a reliable and objective method. In addition to the routine assessment of spe
The aim of these examinations was to check various flow cytometric methods to establish whether they are suitable objective methods for assessing the quality of cryopreserved sperm from stallions. The plasma membrane integrity, the mitochondrial membrane potential, the acrosomal status and the integrity of the chromatin structure in cryopreserved sperm from stallions were assessed. For this purpose various fluorescent stainings were carried out both immediately after the semen samples were thawed and after a three-hour period of incubation at 37 C with SYBR® 14, JC-1 and SYTO® 17 / propidium iodide / FITC-PNA and a sperm chromatin structure analysis (SCSA™) and were then evaluated.
The sperm quality parameters obtained by flow cytometry showed good reproducibility. The results of measurements of three different straws of an ejaculate which were carried out independently of each other did not differ significantly. The intra-class correlation coefficients were between 0.88 and 0.98. The flow cytometric tests evaluated in this examination were therefore considered to be reliable.
In order to demonstrate the correlations between the routine examination methods for assessing the sperm quality and the analysis processes evaluated in this study, the viability and the morphology of the sperms after thawing were assessed by light microscopy using bromide phenol blue nigrosine smears. A computer controlled motility analysis was also carried out. Moderate correlations (0.60 > r > 0.58; p 0.96; p 0.05) with the fertility of the stallions. On the other hand, a negative relationship was established between the integrity of the chromatin structure and the fertility of the stallions (r = - 0.51 / - 0.59).
This study shows that the flow cytometric assessment of the quality of cryopreserved sperm from stallions represents a reliable and objective method. In addition to the routine assessment of spe
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