scholarly journals New Sperm Morphology Analysis in Equids: Trumorph® Vs Eosin-Nigrosin Stain

2021 ◽  
Vol 8 (5) ◽  
pp. 79
Author(s):  
Sabrina Gacem ◽  
Jaime Catalán ◽  
Iván Yánez-Ortiz ◽  
Carles Soler ◽  
Jordi Miró
Keyword(s):  
Phase Contrast ◽  
Morphological Analysis ◽  
Sperm Morphology ◽  
Staining Method ◽  
Good Alternative ◽  
Semen Collection ◽  
Fertility Potential ◽  
Live Sperm ◽  
Subsequent Staining ◽  
Semen Morphology

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 identified using Trumorph®. The use of eosin-nigrosin staining method and Trumorph® led to the same results and both techniques can be used for stallion and donkey sperm morphological analysis. However, considering the fact that Trumorph® uses living sperm helps prevent sperm cell alteration during sample preparation. Therefore, Trumorph® can be a good alternative to the conventional staining method, which provides a quick test on live sperm.

scholarly journals Assessment of stallion semen morphology using two different staining methods, microscopic techniques, and sample sizes

2016 ◽  
Vol 60 (1) ◽  
pp. 99-104 ◽  
Author(s):  
Katarzyna Łącka ◽  
Stanisław Kondracki ◽  
Maria Iwanina ◽  
Anna Wysokińska
Keyword(s):  
Sperm Morphology ◽  
Staining Method ◽  
Microscopic Analysis ◽  
Sample Sizes ◽  
Morphology Analysis ◽  
Morphological Defects ◽  
Staining Methods ◽  
Microscopic Techniques ◽  
Semen Morphology

Abstract Introduction: The aim of this study was to propose the optimal methodology for stallion semen morphology analysis while taking into consideration the staining method, the microscopic techniques, and the workload generated by a number of samples. Material and Methods: Ejaculates from eight pure-bred Arabian horses were tested microscopically for the incidence of morphological defects in the spermatozoa. Two different staining methods (eosin-nigrosin and eosin-gentian dye), two different techniques of microscopic analysis (1000× and 400× magnifications), and two sample sizes (200 and 500 spermatozoa) were used. Results: Well-formed spermatozoa and those with major and minor defects according to Blom’s classification were identified. The applied staining methods gave similar results and could be used in stallion sperm morphology analysis. However, the eosin-nigrosin method was more recommendable, because it allowed to limit the number of visible artefacts without hindering the identification of protoplasm drops and enables the differentiation of living and dead spermatozoa. Conclusion: The applied microscopic techniques proved to be equally efficacious. Therefore, it is practically possible to opt for the simpler and faster 400x technique of analysing sperm morphology to examine stallion semen. We also found that the number of spermatozoa clearly affects the results of sperm morphology evaluation. Reducing the number of spermatozoa from 500 to 200 causes a decrease in the percentage of spermatozoa identified as normal and an increase in the percentage of spermatozoa determined as morphologically defective.

scholarly journals Rotational Dynamics of Optically Trapped Human Spermatozoa

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Elavarasan Subramani ◽  
Himanish Basu ◽  
Shyam Thangaraju ◽  
Sucheta Dandekar ◽  
Deepak Mathur ◽  
...  
Keyword(s):  
Near Infrared ◽  
Beat Frequency ◽  
Translational Motion ◽  
Rotational Dynamics ◽  
Sperm Cells ◽  
Integrated Optical ◽  
Fertility Potential ◽  
Live Sperm ◽  
Fertilization Capacity ◽  
Optically Trapped

Introduction. Optical trapping is a laser-based method for probing the physiological and mechanical properties of cells in a noninvasive manner. As sperm motility is an important criterion for assessing the male fertility potential, this technique is used to study sperm cell motility behavior and rotational dynamics.Methods and Patients. An integrated optical system with near-infrared laser beam has been used to analyze rotational dynamics of live sperm cells from oligozoospermic and asthenozoospermic cases and compared with controls.Results. The linear, translational motion of the sperm is converted into rotational motion on being optically trapped, without causing any adverse effect on spermatozoa. The rotational speed of sperm cells from infertile men is observed to be significantly less as compared to controls.Conclusions. Distinguishing normal and abnormal sperm cells on the basis of beat frequency above 5.6 Hz may be an important step in modern reproductive biology to sort and select good quality spermatozoa. The application of laser-assisted technique in biology has the potential to be a valuable tool for assessment of sperm fertilization capacity for improving assisted reproductive technology.

Standard Sperm Morphology as a Predictor of Male Fertility Potential

1992 ◽  
Vol 28 (1) ◽  
pp. 39-41 ◽  
Author(s):  
J. H. Check ◽  
A. Bollendorf ◽  
M. Press ◽  
T. Blue
Keyword(s):  
Male Fertility ◽  
Sperm Morphology ◽  
Fertility Potential

Quality of domestic cat semen collected by urethral catheterization after the use of different alpha 2-adrenergic agonists

2020 ◽  
pp. 1098612X2097318
Author(s):  
Mónica Madrigal-Valverde ◽  
Rodrigo F Bittencourt ◽  
Antonio DL Ribeiro Filho ◽  
Vivian F Barbosa ◽  
Catharina A Vieira ◽  
...  
Keyword(s):  
Sperm Morphology ◽  
Sperm Concentration ◽  
Domestic Cat ◽  
Domestic Cats ◽  
Adrenergic Agonists ◽  
Urethral Catheterization ◽  
Seminal Parameters ◽  
Semen Collection ◽  
Economical Alternative

Objectives We compared the effects of two alpha (α)2-adrenergic agonists on semen traits. Methods In this study, 13 adult domestic cats were divided into two experimental groups, according to the chemical ejaculation protocol used: the first group received medetomidine hydrochloride (100 µg/kg) and ketamine (5000 µg/kg); the second group received dexmedetomidine hydrochloride (25 µg/kg) and ketamine (5000 µg/kg), both by the intramuscular route. Results The animals responded positively ( P >0.05) to chemical collection. Seminal parameters evaluated included volume, sperm vigor, total motility, progressive motility, sperm concentration, and the structural and functional integrity of the plasma membrane; sperm morphology values did not differ between groups ( P >0.05). Conclusions and relevance The results indicated that dexmedetomidine is a more viable and economical alternative to medetomidine in domestic cats submitted to semen collection by urethral catheterization. Semen collection by urethral catheterization after using α2-adrenergic agonists is a recently developed technique in feline species that is considered to be quick and highly applicable to assisted reproduction programs in felids.

scholarly journals Morphometric dimensions of the stallion sperm head depending on the staining method used

2015 ◽  
Vol 59 (2) ◽  
pp. 263-270 ◽  
Author(s):  
Dorota Banaszewska ◽  
Magdalena Czubaszek ◽  
Renata Walczak-Jędrzejowska ◽  
Katarzyna Andraszek
Keyword(s):  
Surface Area ◽  
Sperm Morphology ◽  
Staining Method ◽  
Sperm Head ◽  
Morphometric Measurements ◽  
Morphological Evaluation ◽  
Length Width ◽  
Staining Techniques ◽  
Control Samples

AbstractSemen was collected from Polish Halfbred stallions. Twenty individuals from 3 to 4 years of age were selected for the study. At least one ejaculate from each stallion was collected and assessed. Sperm morphology was evaluated using Papanicolaou stain, SpermBlue®, and a complex of eosin and gentian stain, whereas unstained slides were used as control samples. Morphometric measurements were performed on 100 randomly selected sperm heads in each ejaculate. The length, width, perimeter, and surface area of the sperm head were measured. The frequency of nuclear vacuoles was determined as well. Tygerberg’s strict criteria, which most precisely characterise the sperm head, were used in the morphological evaluation of the sperm. The results obtained indicate that in the case of staining with SpermBlue®and with eosin+gentian complex, the morphometry of the sperm head seems to be most similar to that observed in unstained smears. It also seems that neither shrinking nor swelling is uniform in the various staining techniques (Papanicolaou stain). Moreover, it appears that in comparison with unstained sperm, all methods caused the width of the head to increase as a result of swelling.

scholarly journals Effect of split ejaculation and seminal extenders on longevity of donkey semen preserved at 5° C

2000 ◽  
Vol 52 (4) ◽  
pp. 372-378 ◽  
Author(s):  
S.L.V. Mello ◽  
M. Henry ◽  
M.C. Souza ◽  
S.M.P. Oliveira
Keyword(s):  
Phase Contrast ◽  
Room Temperature ◽  
Sperm Morphology ◽  
Sperm Concentration ◽  
Egg Yolk ◽  
The Rich ◽  
Student’S T ◽  
The One ◽  
Artificial Vagina ◽  
Student’S T Test

The aim of this study was to evaluate the longevity of donkey sperm comparing the rich seminal fraction and the whole semen in two extenders, Kenney and modified Baken extenders. Semen of five donkeys were collected through an open-end artificial vagina once a week for five consecutive weeks. The two first jets (rich fraction) of semen were collected separately from the rest of the ejaculate. Whole semen samples were obtained mixing proportionally part of the rich with part of the poor seminal fractions. Seminal samples were immediately diluted 1:1 in each extender and maintained at room temperature during sperm concentration analysis. Samples were further diluted to rich 50×10(6) sperm per ml, cooled in a refrigerator at the initial rate of -0.6° C/min and preserved at 5° C. Total motility (TM), progressive motility (PM) and sperm vigor (V) were examined after final dilution and cooling, and every 24 hours up to the decrease of total motility under 10%. Sperm morphology was evaluated using a phase contrast microscope directly after dilution, on days 3, 6 and 9 post collection. It was used a 2×2 factorial design in a randomised bloc experiment, and means were compared by Student’s t test. Longevity did not vary between the rich seminal fraction and the whole semen for both extenders used. TM, PM, V and sperm morphology were better preserved in the extender with egg yolk (modified Baken extender) than in the one with skimmed milk (Kenney) in both seminal fractions.

Direct three-dimensional imaging for morphological analysis of electrospun fibers with laboratory-based Zernike X-ray phase-contrast computed tomography

2020 ◽  
Vol 115 ◽  
pp. 111045 ◽  
Author(s):  
Cristine Santos de Oliveira ◽  
Adriana Trompetero González ◽  
Tobias Hedtke ◽  
Tobias Kürbitz ◽  
Andreas Heilmann ◽  
...  
Keyword(s):  
Computed Tomography ◽  
Phase Contrast ◽  
Morphological Analysis ◽  
Three Dimensional ◽  
Electrospun Fibers ◽  
Three Dimensional Imaging ◽  
X Ray

A new technique for analysis of human sperm morphology in unstained cells from raw semen

2016 ◽  
Vol 28 (4) ◽  
pp. 428 ◽  
Author(s):  
Carles Soler ◽  
Almudena García-Molina ◽  
María Sancho ◽  
Jesús Contell ◽  
Manuel Núñez ◽  
...  
Keyword(s):  
Phase Contrast ◽  
Sperm Morphology ◽  
Semen Analysis ◽  
Human Sperm ◽  
New Method ◽  
Phase Contrast Microscopy ◽  
Robust Method ◽  
Morphology Analysis ◽  
Contrast Microscopy ◽  
A New Technique

Sperm morphology analysis is a fundamental component of semen analysis, but its real significance has been clouded by the plethora of techniques used for its evaluation. Most involve different fixation and staining procedures that induce artefacts. Herein we describe Trumorph (Proiser R+D, Paterna, Spain), a new method for sperm morphology analysis based on examination of wet preparations of spermatozoa immobilised, after a short 60°C shock, in narrow chambers and examined by negative phase contrast microscopy. A range of morphological forms was observed, similar to those found using conventional fixed and stained preparations, but other forms were also found, distinguishable only by the optics used. The ease of preparation makes the Trumorph a robust method applicable for the analysis of living unmodified spermatozoa in a range of situations. Subsequent studies on well-characterised samples are required to describe the morphology of spermatozoa with fertilising potential.

Molecular biology of sperm maturation in the human epididymis

1995 ◽  
Vol 4 (3) ◽  
pp. 121-139 ◽  
Author(s):  
Christiane Kirchhoff
Keyword(s):  
Molecular Biology ◽  
Male Infertility ◽  
Sperm Morphology ◽  
Basic Research ◽  
Laboratory Animals ◽  
Oxygen Species ◽  
Autoimmune Reaction ◽  
Human Epididymis ◽  
Fertility Potential ◽  
Sperm Movement

Male infertility is a frequent cause of childlessness, and, indeed, a comparison of the contributions to conception failure made by male and female factors shows them to be equally frequent. In practice, male infertility appears to be resistant to most treatments. However, the major reason for this may be that often attempts are carried out without knowing the cause of the problem. Unlike in women, obstructions and hormonal disorders are rare in male infertility. Rather, it would appear that sperm disorders are the most common cause, reflecting a variety of pathogenetic mechanisms. Defects in sperm morphology, defective sperm movement, deficient development or functional failure of the acrosome, and the excessive generation of reactive oxygen species are changes that are often seen in infertile semen, but little is known about their aetiology. In 5–10% of men being treated for infertilty, an autoimmune reaction against spermatozoa is observed. Although the correlation between the presence of systemic antisperm antibodies and fertility potential is poor, the appearance of sperm-bound antibodies of immunoglobulin class IgA in semen seems to be closely associated with infertility. Studies in laboratory animals and humans have shown that complementary adhesion molecules are located on the surface of oocytes and spermatozoa. These molecules interact and lead to gamete fusion. Abnormalities in these molecules on the sperm surface might be expected to contribute to male infertility. However, their clinical significance has not yet been documented, and the molecular basis of human gamete interaction is far from being understood. Therefore, the key to understanding male infertility may lie in basic research which directly targets the fundamental cellular and molecular biology of the human spermatozoon.

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