Carles Soler - Academia.edu (original) (raw)

Papers by Carles Soler

Revista Internacional de Andrología

Biology, 2021

The aim was to determine the relationship between kinematic parameters of boar spermatozoa and fe... more The aim was to determine the relationship between kinematic parameters of boar spermatozoa and fertility rates of sow, as well as to assess the effect of sperm clusters on the fertility capacity of the ejaculate. Semen samples were collected from 11 sexually mature boars. Samples were analyzed by an ISAS®v1 CASA-Mot system for eight kinematic parameters. Ejaculate clusters were characterized using multivariate procedures, such as principal factors (PFs) analysis and clustering methods (the k-means model). Four different ejaculate clusters were identified from two kinematic PFs which involved linear trajectory and velocity. There were differences (p < 0.05) between the sperm kinematic variables by sire line. There was no statistical difference (p > 0.05) between dam lines and ejaculate clusters in fertility variables. The discriminant ability of the different kinematics of sperm variables to predict litter size fertility was analyzed using receiver operating characteristics (RO...

South American camelids are classified in four different species included in two genera. Llama (L... more South American camelids are classified in four different species included in two genera. Llama (Lama glama L.) and alpaca (Vicunya pacos, L.) are the domestic species, presenting a high economical interest. This classification is based on historical, evolutionary and morphological criteria, but the level of hybridization and the total reproductive compatibility make it difficult to use the “biological criteria” based on reproductive isolation. The aim of the work was to see if there are differences on spermatozoa morphometry indicating a pre-zygotic isolation mechanism based on sperm competition. Nine adult Lanuda (Ch’aku) llamas and eleven Huacaya alpacas were used. After obtaining the samples using an artificial vagina, semen smears were stained with Hemacolor kit. Samples were automatically analysed using the morphometry module of the ISAS ® v1 CASA system. Almost 100 sperm cells were analysed per sample. The following parameters were calculated: Length, Width; Perimeter, Area, E...

Veterinary Sciences, 2021

The evaluation of the male fertility potential is based on the analysis of the basic spermatic ch... more The evaluation of the male fertility potential is based on the analysis of the basic spermatic characteristics of concentration, motility and morphology. Thus, the study of sperm morphology is a fundamental element in the seminal analysis, but its real meaning has been biased by the techniques used for its evaluation. These techniques involve dehydration phases and subsequent staining, which involves the production of artifacts. The aim of the study is to compare two methods for equid semen morphology evaluation, Trumorph® using living sperm vs. eosin-nigrosine stain. A total of 49 ejaculates from stallions and donkeys were used. After semen collection and dilution, an aliquot was placed on the slide and introduced in the Trumorph® device. Then observation was made with a 40x objective and negative phase-contrast microscope. Another aliquot was stained using eosin-nigrosine stain and viewed using 100× magnification. Well-formed sperm were observed, and different abnormalities were i...

Animal Reproduction Science, 2020

This study was conducted to determine optimum image capture frame rates (FRO) when there was eval... more This study was conducted to determine optimum image capture frame rates (FRO) when there was evaluation of different types of counting chambers used for CASA-Mot determinations of stallion sperm motility. Sperm VCL was determined at frame rates of 25-250 f/s in: 1) Sperm-track® (Spk) 10 and 20 chambers (drop displacement-type chambers 10 and 20 μm-deep respectively; and 2) ISAS®D4C10, ISAS®D4C20 (10 and 20 μm-deep respectively) and ISAS®D4C20 L (20 μm-deep) capillary loaded chambers. Values for different sperm kinematic variables were determined using each chamber at 250 f/s, which is the maximum frame rate that the software can be used for analyses. With evaluation of Spk chambers, there was a greater curvilinear velocity (VCL), average path velocity (VAP), straight line velocity (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) values (P < 0.05) than with capillary loaded chambers, with there being greatest values with 20 μm-deep chambers. With the Spk10 chamber, VCL and ALH were greater at the chamber centre than periphery. There were no such differences for the Spk20 chamber. With evaluation of the D4C10 chamber, VSL and STR were less when there was a sperm deposition point towards the chamber end, while there were the opposite for the D4C20 chamber. When there was evaluation of the D4C20 chamber, there were also greater VCL, WOB and BCF values in distal areas. With use of most of these chambers, data should be collected from different fields and means determined, however, this is not necessary with Spk20 chambers.

Revista de Investigaciones Veterinarias del Perú, 2020

El objetivo del estudio fue determinar el efecto del tratamiento con papaína en la cinética de lo... more El objetivo del estudio fue determinar el efecto del tratamiento con papaína en la cinética de los espermatozoides de llama. Se seleccionaron 4 llamas machos Ch’aku de 5-8 años de edad. El semen fue colectado mediante vagina artificial y maniquí. Se hicieron cuatro colectas de semen por animal con intervalos de una semana. Las muestras de semen fueron separadas en dos alícuotas. Una para su análisis en fresco y la otra para ser tratada con papaína (20 minutos). Se evaluó el volumen, filancia, concentración de espermatozoides, porcentaje de espermatozoides vivos, porcentaje de funcionalidad de la membrana espermática, y porcentaje de espermatozoides con reacción acrosomal. La movilidad de los espermatozoides y su cinética se evaluó utilizando un sistema CASA-Mot (ISAS®), determinándose la movilidad total (MT) y la movilidad progresiva (MP), así como los parámetros de cinética espermática: velocidad curvilínea (VCL), velocidad rectilínea (VSL), velocidad promedio (VAP), y los índices ...

Frontiers in Veterinary Science, 2021

This study aimed to characterize the sperm kinematic values with high frames per second, to defin... more This study aimed to characterize the sperm kinematic values with high frames per second, to define the subpopulation structure of a horse and a donkey and compare them. A total of 57 fresh semen ejaculates (26 Spanish and 16 Arabian horse breeds and 10 donkeys) were collected and subsequently analyzed for kinematic parameters using the Computer-aided sperm motility analysis ISAS®v1.2 system and using a Spermtrack® 10-μm depth counting chamber. Sequences were recorded at 250 frames per second, and eight kinematic parameters were automatically evaluated. All kinematic parameters showed significant differences between a donkey and a horse and between horse breeds. All ejaculates evaluated showed excellent semen motility characteristics, with significantly higher values for all kinematic parameters for donkeys compared with horses except for beat-cross frequency. Donkey sperm was faster and linear than the horse. Regarding horse breeds differences, the Spanish horse had higher average p...

Animals, 2021

The aim of the study was to compare the morphometric features of sperm head size and shape from t... more The aim of the study was to compare the morphometric features of sperm head size and shape from the Pietrain line and the Duroc × Pietrain boar crossbred terminal lines, and to evaluate their relationship with reproductive success after artificial insemination of sows produced from crossbreeding the York, Landrace and Pietrain breeds. Semen samples were collected from 11 sexually mature boars. Only ejaculates with greater than 70% motility rate and <15% of abnormal sperm were used for artificial inseminations (AI) and included in the study. Samples were analyzed using an ISAS®v1 computer-assisted sperm analysis system for eight morphometric parameters of head shape and size (CASA-Morph). Sub-populations of morphometric ejaculates were characterized using multivariate procedures, such as principal component (PC) analysis and clustering methods (k-means model). Four different ejaculate sub-populations were identified from two PCs that involved the head shape and size of the spermat...

Animal Reproduction Science, 2020

There has been very limited use of computer assisted semen analysis (CASA) to evaluate reptile sp... more There has been very limited use of computer assisted semen analysis (CASA) to evaluate reptile sperm. The aim of this study was to examine sperm kinematic variables in American crocodile (Crocodylus acutus) semen samples and to assess whether sperm subpopulations could be characterized. Eight ejaculates (two ejaculates/male) from four sexually mature captive crocodiles were obtained. An ISAS®v1 CASA-Mot system, with an image acquisition rate of 50 Hz, and ISAS®D4C20 counting chambers were used for sperm analyses. The percentages of motile and progressively motile spermatozoa did not differ among animals (P > 0.05) but there was a significant animal effect with regards to kinematic variables (P < 0.05). Principal component (PC) analysis revealed that kinematic variables grouped into three components: PC1, related to velocity; PC2 to progressiveness and PC3 to oscillation. Subpopulation structure analysis identified four groups (P < 0.05), which represented, on average, 9.8%, 32.1%, 26.8%, and 31.3% of the total sperm population. Males differed in the proportion of sperm in each of the kinematic subpopulations. This new approach for the analysis of reptile sperm kinematic subpopulations, reflecting quantifiable parameters generated by CASA system technology, opens up possibilities for future assessments of crocodile sperm and will be useful in the future development of assisted reproduction for these species.

Animals, 2020

In order to optimize the donkey sperm motility analysis by the CASA (Computer Assisted Sperm Anal... more In order to optimize the donkey sperm motility analysis by the CASA (Computer Assisted Sperm Analysis)-Mot system, twelve ejaculates were collected from six jackasses. Capillary loaded chamber (CLC), ISAS®D4C depths 10 and 20 µm, ISAS®D4C Leja 20 and drop displacement chamber (DDC), Spermtrack® (Spk) depths 10 and 20 µm were used. Sperm kinematic variables were evaluated using each chamber and a high-resolution camera capable of capturing a maximum of 500 frames/second (fps). The optimum frame rate (OFR) (defined according to curvilinear velocity—VCL) was dependent on chamber type. The highest OFR obtained was 278.46 fps by Spk20. Values for VCL, straight-line velocity (VSL), straightness (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) were high in DDC and 10 µm depth. In both DDC 10 and 20 µm, the sperm velocities (VCL, VSL, VAP) and ALH values decreased significantly from the centre to the edges, while Wobble and BCF increased. No defined behavio...

Biology, 2020

The ejaculate is heterogenous and sperm sub-populations with different kinematic patterns can be ... more The ejaculate is heterogenous and sperm sub-populations with different kinematic patterns can be identified in various species. Nevertheless, although these sub-populations are statistically well defined, the statistical differences are not always relevant. The aim of the present study was to characterize kinematic sub-populations in sperm from two bovine species, and diluted with different commercial extenders, and to determine the statistical relevance of sub-populations through Bayesian analysis. Semen from 10 bulls was evaluated after thawing. An ISAS®v1 computer-assisted sperm analysis (CASA)-Mot system was employed with an image acquisition rate of 50 Hz and ISAS®D4C20 counting chambers. Sub-populations of motile spermatozoa were characterized using multivariate procedures such as principal components (PCs) analysis and clustering methods (k-means model). Four different sperm sub-populations were identified from three PCs that involved progressiveness, velocity, and cell undul...

Asian Journal of Andrology, 2020

MATERIALS AND METHODS Semen collection and processing All animals were handled according to the p... more MATERIALS AND METHODS Semen collection and processing All animals were handled according to the procedures approved by the University of Zaragoza Ethics Committee (Zaragoza, Spain), and the research was performed in accordance with the Spanish Policy for Animal Protection (RD53/2013), which conforms to European Union Directive 86/609 regarding the protection of animals used in scientific experiments. The study analyzed cryopreserved semen samples collected from 16 commercial Holstein bulls. The ejaculate was then extended in BullXcell (IMV Technologies, Humeco, Huesca, Spain) to a final concentration of 23 × 10 6 sperm per 0.25 ml semen straw (IMV Technologies). Straws were cooled to 4°C over 3 h and then frozen to −140°C as follows: −5°C per min from +4°C to −10°C, −40°C

Animal Reproduction Science, 2019

Animal Reproduction Science, 2019

Kinematic and head morphometric characterisation of spermatozoa from the Brown Caiman (Caiman cro... more Kinematic and head morphometric characterisation of spermatozoa from the Brown Caiman (Caiman crocodilus fuscus)

Asian Journal of Andrology, 2019

Reproduction in Domestic Animals, 2019

Veterinární Medicína, 2012

Computer-assisted sperm morphometry analysis has improved the assessment of sperm morphology, but... more 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 ...

Revista Internacional de Andrología

Biology, 2021

The aim was to determine the relationship between kinematic parameters of boar spermatozoa and fe... more The aim was to determine the relationship between kinematic parameters of boar spermatozoa and fertility rates of sow, as well as to assess the effect of sperm clusters on the fertility capacity of the ejaculate. Semen samples were collected from 11 sexually mature boars. Samples were analyzed by an ISAS®v1 CASA-Mot system for eight kinematic parameters. Ejaculate clusters were characterized using multivariate procedures, such as principal factors (PFs) analysis and clustering methods (the k-means model). Four different ejaculate clusters were identified from two kinematic PFs which involved linear trajectory and velocity. There were differences (p < 0.05) between the sperm kinematic variables by sire line. There was no statistical difference (p > 0.05) between dam lines and ejaculate clusters in fertility variables. The discriminant ability of the different kinematics of sperm variables to predict litter size fertility was analyzed using receiver operating characteristics (RO...

South American camelids are classified in four different species included in two genera. Llama (L... more South American camelids are classified in four different species included in two genera. Llama (Lama glama L.) and alpaca (Vicunya pacos, L.) are the domestic species, presenting a high economical interest. This classification is based on historical, evolutionary and morphological criteria, but the level of hybridization and the total reproductive compatibility make it difficult to use the “biological criteria” based on reproductive isolation. The aim of the work was to see if there are differences on spermatozoa morphometry indicating a pre-zygotic isolation mechanism based on sperm competition. Nine adult Lanuda (Ch’aku) llamas and eleven Huacaya alpacas were used. After obtaining the samples using an artificial vagina, semen smears were stained with Hemacolor kit. Samples were automatically analysed using the morphometry module of the ISAS ® v1 CASA system. Almost 100 sperm cells were analysed per sample. The following parameters were calculated: Length, Width; Perimeter, Area, E...

Veterinary Sciences, 2021

The evaluation of the male fertility potential is based on the analysis of the basic spermatic ch... more The evaluation of the male fertility potential is based on the analysis of the basic spermatic characteristics of concentration, motility and morphology. Thus, the study of sperm morphology is a fundamental element in the seminal analysis, but its real meaning has been biased by the techniques used for its evaluation. These techniques involve dehydration phases and subsequent staining, which involves the production of artifacts. The aim of the study is to compare two methods for equid semen morphology evaluation, Trumorph® using living sperm vs. eosin-nigrosine stain. A total of 49 ejaculates from stallions and donkeys were used. After semen collection and dilution, an aliquot was placed on the slide and introduced in the Trumorph® device. Then observation was made with a 40x objective and negative phase-contrast microscope. Another aliquot was stained using eosin-nigrosine stain and viewed using 100× magnification. Well-formed sperm were observed, and different abnormalities were i...

Animal Reproduction Science, 2020

This study was conducted to determine optimum image capture frame rates (FRO) when there was eval... more This study was conducted to determine optimum image capture frame rates (FRO) when there was evaluation of different types of counting chambers used for CASA-Mot determinations of stallion sperm motility. Sperm VCL was determined at frame rates of 25-250 f/s in: 1) Sperm-track® (Spk) 10 and 20 chambers (drop displacement-type chambers 10 and 20 μm-deep respectively; and 2) ISAS®D4C10, ISAS®D4C20 (10 and 20 μm-deep respectively) and ISAS®D4C20 L (20 μm-deep) capillary loaded chambers. Values for different sperm kinematic variables were determined using each chamber at 250 f/s, which is the maximum frame rate that the software can be used for analyses. With evaluation of Spk chambers, there was a greater curvilinear velocity (VCL), average path velocity (VAP), straight line velocity (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) values (P < 0.05) than with capillary loaded chambers, with there being greatest values with 20 μm-deep chambers. With the Spk10 chamber, VCL and ALH were greater at the chamber centre than periphery. There were no such differences for the Spk20 chamber. With evaluation of the D4C10 chamber, VSL and STR were less when there was a sperm deposition point towards the chamber end, while there were the opposite for the D4C20 chamber. When there was evaluation of the D4C20 chamber, there were also greater VCL, WOB and BCF values in distal areas. With use of most of these chambers, data should be collected from different fields and means determined, however, this is not necessary with Spk20 chambers.

Revista de Investigaciones Veterinarias del Perú, 2020

El objetivo del estudio fue determinar el efecto del tratamiento con papaína en la cinética de lo... more El objetivo del estudio fue determinar el efecto del tratamiento con papaína en la cinética de los espermatozoides de llama. Se seleccionaron 4 llamas machos Ch’aku de 5-8 años de edad. El semen fue colectado mediante vagina artificial y maniquí. Se hicieron cuatro colectas de semen por animal con intervalos de una semana. Las muestras de semen fueron separadas en dos alícuotas. Una para su análisis en fresco y la otra para ser tratada con papaína (20 minutos). Se evaluó el volumen, filancia, concentración de espermatozoides, porcentaje de espermatozoides vivos, porcentaje de funcionalidad de la membrana espermática, y porcentaje de espermatozoides con reacción acrosomal. La movilidad de los espermatozoides y su cinética se evaluó utilizando un sistema CASA-Mot (ISAS®), determinándose la movilidad total (MT) y la movilidad progresiva (MP), así como los parámetros de cinética espermática: velocidad curvilínea (VCL), velocidad rectilínea (VSL), velocidad promedio (VAP), y los índices ...

Frontiers in Veterinary Science, 2021

This study aimed to characterize the sperm kinematic values with high frames per second, to defin... more This study aimed to characterize the sperm kinematic values with high frames per second, to define the subpopulation structure of a horse and a donkey and compare them. A total of 57 fresh semen ejaculates (26 Spanish and 16 Arabian horse breeds and 10 donkeys) were collected and subsequently analyzed for kinematic parameters using the Computer-aided sperm motility analysis ISAS®v1.2 system and using a Spermtrack® 10-μm depth counting chamber. Sequences were recorded at 250 frames per second, and eight kinematic parameters were automatically evaluated. All kinematic parameters showed significant differences between a donkey and a horse and between horse breeds. All ejaculates evaluated showed excellent semen motility characteristics, with significantly higher values for all kinematic parameters for donkeys compared with horses except for beat-cross frequency. Donkey sperm was faster and linear than the horse. Regarding horse breeds differences, the Spanish horse had higher average p...

Animals, 2021

The aim of the study was to compare the morphometric features of sperm head size and shape from t... more The aim of the study was to compare the morphometric features of sperm head size and shape from the Pietrain line and the Duroc × Pietrain boar crossbred terminal lines, and to evaluate their relationship with reproductive success after artificial insemination of sows produced from crossbreeding the York, Landrace and Pietrain breeds. Semen samples were collected from 11 sexually mature boars. Only ejaculates with greater than 70% motility rate and <15% of abnormal sperm were used for artificial inseminations (AI) and included in the study. Samples were analyzed using an ISAS®v1 computer-assisted sperm analysis system for eight morphometric parameters of head shape and size (CASA-Morph). Sub-populations of morphometric ejaculates were characterized using multivariate procedures, such as principal component (PC) analysis and clustering methods (k-means model). Four different ejaculate sub-populations were identified from two PCs that involved the head shape and size of the spermat...

Animal Reproduction Science, 2020

There has been very limited use of computer assisted semen analysis (CASA) to evaluate reptile sp... more There has been very limited use of computer assisted semen analysis (CASA) to evaluate reptile sperm. The aim of this study was to examine sperm kinematic variables in American crocodile (Crocodylus acutus) semen samples and to assess whether sperm subpopulations could be characterized. Eight ejaculates (two ejaculates/male) from four sexually mature captive crocodiles were obtained. An ISAS®v1 CASA-Mot system, with an image acquisition rate of 50 Hz, and ISAS®D4C20 counting chambers were used for sperm analyses. The percentages of motile and progressively motile spermatozoa did not differ among animals (P > 0.05) but there was a significant animal effect with regards to kinematic variables (P < 0.05). Principal component (PC) analysis revealed that kinematic variables grouped into three components: PC1, related to velocity; PC2 to progressiveness and PC3 to oscillation. Subpopulation structure analysis identified four groups (P < 0.05), which represented, on average, 9.8%, 32.1%, 26.8%, and 31.3% of the total sperm population. Males differed in the proportion of sperm in each of the kinematic subpopulations. This new approach for the analysis of reptile sperm kinematic subpopulations, reflecting quantifiable parameters generated by CASA system technology, opens up possibilities for future assessments of crocodile sperm and will be useful in the future development of assisted reproduction for these species.

Animals, 2020

In order to optimize the donkey sperm motility analysis by the CASA (Computer Assisted Sperm Anal... more In order to optimize the donkey sperm motility analysis by the CASA (Computer Assisted Sperm Analysis)-Mot system, twelve ejaculates were collected from six jackasses. Capillary loaded chamber (CLC), ISAS®D4C depths 10 and 20 µm, ISAS®D4C Leja 20 and drop displacement chamber (DDC), Spermtrack® (Spk) depths 10 and 20 µm were used. Sperm kinematic variables were evaluated using each chamber and a high-resolution camera capable of capturing a maximum of 500 frames/second (fps). The optimum frame rate (OFR) (defined according to curvilinear velocity—VCL) was dependent on chamber type. The highest OFR obtained was 278.46 fps by Spk20. Values for VCL, straight-line velocity (VSL), straightness (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) were high in DDC and 10 µm depth. In both DDC 10 and 20 µm, the sperm velocities (VCL, VSL, VAP) and ALH values decreased significantly from the centre to the edges, while Wobble and BCF increased. No defined behavio...

Biology, 2020

The ejaculate is heterogenous and sperm sub-populations with different kinematic patterns can be ... more The ejaculate is heterogenous and sperm sub-populations with different kinematic patterns can be identified in various species. Nevertheless, although these sub-populations are statistically well defined, the statistical differences are not always relevant. The aim of the present study was to characterize kinematic sub-populations in sperm from two bovine species, and diluted with different commercial extenders, and to determine the statistical relevance of sub-populations through Bayesian analysis. Semen from 10 bulls was evaluated after thawing. An ISAS®v1 computer-assisted sperm analysis (CASA)-Mot system was employed with an image acquisition rate of 50 Hz and ISAS®D4C20 counting chambers. Sub-populations of motile spermatozoa were characterized using multivariate procedures such as principal components (PCs) analysis and clustering methods (k-means model). Four different sperm sub-populations were identified from three PCs that involved progressiveness, velocity, and cell undul...

Asian Journal of Andrology, 2020

MATERIALS AND METHODS Semen collection and processing All animals were handled according to the p... more MATERIALS AND METHODS Semen collection and processing All animals were handled according to the procedures approved by the University of Zaragoza Ethics Committee (Zaragoza, Spain), and the research was performed in accordance with the Spanish Policy for Animal Protection (RD53/2013), which conforms to European Union Directive 86/609 regarding the protection of animals used in scientific experiments. The study analyzed cryopreserved semen samples collected from 16 commercial Holstein bulls. The ejaculate was then extended in BullXcell (IMV Technologies, Humeco, Huesca, Spain) to a final concentration of 23 × 10 6 sperm per 0.25 ml semen straw (IMV Technologies). Straws were cooled to 4°C over 3 h and then frozen to −140°C as follows: −5°C per min from +4°C to −10°C, −40°C

Animal Reproduction Science, 2019

Animal Reproduction Science, 2019

Kinematic and head morphometric characterisation of spermatozoa from the Brown Caiman (Caiman cro... more Kinematic and head morphometric characterisation of spermatozoa from the Brown Caiman (Caiman crocodilus fuscus)

Asian Journal of Andrology, 2019

Reproduction in Domestic Animals, 2019

Veterinární Medicína, 2012

Computer-assisted sperm morphometry analysis has improved the assessment of sperm morphology, but... more 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 ...