Computer automated sperm head morphometry analysis (ASMA) of goat spermatozoa

Gravance, Curtis G.; Lewis, Kenneth; Casey, Patrick J.

doi:10.1016/0093-691x(95)00286-h

Cited by 47 publications

(57 citation statements)

References 12 publications

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“…However, the results indicate that 100 properly digitised sperm cells appeared sufficient for the morphometric characterization of a stallion semen sample under these conditions, as it produced similar measurements as analysing 150, 175 or 200 spermatozoa. The analysis of 100 spermatozoa obtain accurate measurements and greatly reduces the time to perform an analysis, which was in agreement with results obtained in dog (Rijsselaere et al, 2004) and goat (Gravance et al, 1995). However, because heterogeneous abnormal sperm head morphology of equine species had been described previously, it is possible that to overcome possible problems associated with the evaluation of infertile samples, in these semen specimens a high number of spermatozoa should be analysed.…”

Section: Discussionsupporting

confidence: 62%

“…The performance of the SCA has been evaluated using three staining methods. Among the methods tested for stallions, Trypan blue and Giemsa (Kusunoki et al, 1988), Papanicolau (Hafez, 1987) and Spermac (Oettle, 1986) are not suitable for ASMA systems as they result in poorly stained cells, which do not permit digitisation (Gravance et al, 1995). The staining methods compared in this study (DQ, HC, HH) were chosen based on the positive results obtained in humans and in several animal species for different ASMA systems (Lacquet et al, 1996;Gago et al, 1998;Soler et al, 2000).…”

Section: Discussionmentioning

confidence: 99%

“…Currently, the precision of ASMA systems depends upon the standardization of these variables Gago et al, 1998;Gravance et al, 1995;Hidalgo et al, 2004), namely appropriate sample preparation (washing, fixation and staining) and correct microscopic image analysis. In addition to the variations inherent to the evaluation process, errors are often the result of differences between ASMA systems or the fact that an insufficient number of spermatozoa are analysed which are not representative of the sample.…”

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confidence: 99%

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Effect of sample size and staining methods on stallion sperm morphometry by the Sperm Class Analyzer

Hidalgo¹,

Rodríguez²,

Dorado³

et al. 2005

Vet. Med.

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ABSTRACT:Computer-assisted sperm morphometry analysis has improved the assessment of sperm morphology, but the results depend on the use of adequate evaluation and staining procedures of spermatozoa from individual species. In this study, the morphological module of the Sperm Class Analyzer ® was used for the morphometric analysis of stallion sperm heads and midpieces. Semen samples were obtained from six fertile stallions in order to evaluate the influence of three staining procedures (Diff-Quik, Hemacolor and Harris' Haematoxylin) on the accuracy of image processing and sperm morphometry, and the effect of the sample size on sperm morphometric measurements. Harris' Haematoxylin was the staining technique of choice on the accuracy of the image processing with an optimum contrast of sperm cells with the surrounding background that allows an efficient boundary detection and segmentation which results in the highest proportion of sperm heads and midpieces assessed (80.47%). The results indicate that the staining methods affected significantly the sperm dimensions with increased values from Diff-Quik than Hemacolor and Harris' Haematoxylin respectively (Diff-Quik > Hemacolor > Harris' Haematoxylin). No differences in morphometric parameters were found when 100, 150, 175 or 200 spermatozoa were analysed. In conclusion, to obtain objective and accurate sperm morphometric measurements by the Sperm Class Analyzer ® system in the stallion, it's recommended the analysis of 100 spermatozoa from slides which have been previously stained with Harris' Haematoxylin.

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Section: Discussionsupporting

confidence: 62%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Effect of sample size and staining methods on stallion sperm morphometry by the Sperm Class Analyzer

Hidalgo¹,

Rodríguez²,

Dorado³

et al. 2005

Vet. Med.

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“…The processes utilized for cooled and frozen storage of sperm may cause cellular damage that could compound both of these factors. Conventional measures of semen quality, which include compensable variables such as motility and morphology, are measures of the overall population of sperm and indicate gross functionality (Gravance and Davis, 1995). These assessments are routinely performed, but the ability to predict fertility based on the amount of morphologically abnormal sperm has varied among authors (Torres-BogThis project was supported by the Center for Equine Health with funds provided by the Oak Tree Racing Association, the State of California pari-mutuel fund, and contributions by private donors.…”

mentioning

confidence: 99%

Detection of DNA Damage in Response to Cooling Injury in Equine Spermatozoa Using Single‐Cell Gel Electrophoresis

Linfor

Meyers

2002

Journal of Andrology

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Single-cell gel electrophoresis (SCGE), or comet assay, has the ability to detect damage at the single cell level and has not been reported for equine sperm. The ability to detect nuclear damage at the single cell level could aid in the advancement of protocols for optimal semen preservation. The goals of these experiments were to adapt this assay for use with equine sperm and to utilize the assay for determining the integrity of equine sperm DNA following treatments with storage at various decreased temperatures (Ϫ20ЊC and 5ЊC). Results from experiments in which sperm were frozen (Ϫ20ЊC) in the absence of cryoprotectants revealed that significantly more cells with fragmented tails of DNA, or comets, occurred among those exposed to 1, 3, and 5 freeze-thaw cycles (65% Ϯ 6%, 76% Ϯ 11%, 92% Ϯ 6%, respectively) compared with fresh, untreated sperm (19% Ϯ 16%, P Ͻ .05). In addition, DNA damage was different (P Ͻ .05) between the three freeze-thaw treatments. Sensitivity of SCGE on equine sperm was further tested with known ratios of frozen-thawed and fresh cells. The amount of detectable DNA damage was positively correlated with the percentage of cryodamaged cells in each treatment (r 2 ϭ 0.92, P Ͻ .05). Potential damage as a result of cooled storage was also investigated and results revealed that sperm stored for 48 hours (at 5ЊC) had a higher percentage of comets than that of fresh sperm (63% Ϯ 13.9% and 28% Ϯ 15.6%, respectively, P Ͻ .05). The percentage of viable sperm also decreased linearly over time and was inversely correlated with percent of comets (r 2 ϭ 0.805, P Ͻ .001). Detection of sublethal and/or uncompensable fertility factors in semen, such as DNA fragmentation, could be useful for detecting male differences in semen for cooling or cryopreservation potential and could provide a tool for monitoring and preserving fertility for individual stallions.

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“…Understanding the specific role of biophysical factors is important for fertility investigations and for improving the success of assisted reproductive technologies. Because of that, different analysis methods have been developed, as the morphometric sperm head analysis (ASMA) includes a computerized analysis system that provides a series of accurate, objective and repeatable parameters that have been reported for several different species such as caprine, ovine, porcine, rabbit, equine, non-human primates and bovine species (Davis et al, 1993;Gravance and Davis, 1995;Garcia-Herreros et al, 2006;Marti et al, 2011a;Valle et al, 2012;Garcia-Herreros et al, 2014a). Development of the ASMA system enables the evaluation of new sperm characteristics that until now have not been possible, such as sperm nuclear shape assessment using Fourier functions and harmonic amplitudes (Ostermeier et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

Sperm volumetric dynamics during in vitro capacitation process in bovine spermatozoa

García-Herreros

Leal

2015

Animal

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Previous studies have demonstrated that sperm head morphometry can be used as a potential diagnostic tool for detecting biophysical changes associated with sperm viability in bovine spermatozoa. In this study, sperm head morphometry was used to investigate its value as a biophysical marker for detecting volumetric changes in bovine spermatozoa under in vitro capacitating and non-capacitating incubation conditions. To further test this hypotesis, aliquots of pooled, washed bovine sperm were incubated in either Tyrode's complete medium with heparin (TCMH; a capacitating medium containing Ca 2+ , NaHCO 3 and heparin), Tyrode's complete medium heparin-free (TCM; a medium containing just Ca 2+ and NaHCO 3 ) or Tyrode's basal medium (TBM; a non-capacitating medium free of Ca 2+ , NaHCO 3 and heparin, used as control). Aliquots of sperm were processed for morphometric analysis at different incubation-time intervals (0, 3 and 6 h at 38°C), and the chlortetracycline assay was used simultaneously to confirm the ability of the sperm to undergo capacitation (B pattern) and the acrosome reaction (AR pattern) status in each medium. After 3 h of incubation under TCMH conditions, a significant increase was observed in the percentage of B and AR patterns and a significant decrease was found in all sperm morphometric parameters ( P < 0.01). Interestingly, after 6 h of incubation in TCMH, the percentage of B and AR patterns increased drastically over time and marked differences were found in the dimensional and shape parameters, which were significantly smaller compared with TBM or TCM media ( P < 0.001). Significant correlations were observed between sperm size and AR pattern (r = −0.875, P < 0.01). In conclusion, sperm head morphometry can be used as a potential biophysical marker for detecting volumetric changes during capacitation process in bovine spermatozoa.

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Computer automated sperm head morphometry analysis (ASMA) of goat spermatozoa

Cited by 47 publications

References 12 publications

Effect of sample size and staining methods on stallion sperm morphometry by the Sperm Class Analyzer

Effect of sample size and staining methods on stallion sperm morphometry by the Sperm Class Analyzer

Detection of DNA Damage in Response to Cooling Injury in Equine Spermatozoa Using Single‐Cell Gel Electrophoresis

Sperm volumetric dynamics during in vitro capacitation process in bovine spermatozoa

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