Scanning electron microscopy of human sperms after preparation of semen for in-vitro fertilization

Grab, Dieter; Thierauf, S.; Rosenbusch, B.; Sterzik, K.

doi:10.1007/bf02456677

Cited by 5 publications

(2 citation statements)

References 6 publications

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“…Conventional TEM was instrumental in the prognosis of male fertility by identifying teratozoospermia [42,43]. Likewise, SEM has been applied extensively to assess the detailed surface morphologies of normal and abnormal spermatozoa of humans and other mammalian species [28,44,45]. SEM can also be used as a tool to evaluate and score morphological abnormalities of spermatozoa in infertile men [46].…”

Section: Conventional Em Has Contributed Greatly To Our Understandingmentioning

confidence: 99%

Looking back and looking forward: contributions of electron microscopy to the structural cell biology of gametes and fertilization

2020

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Mammalian gametes—the sperm and the egg—represent opposite extremes of cellular organization and scale. Studying the ultrastructure of gametes is crucial to understanding their interactions, and how to manipulate them in order to either encourage or prevent their union. Here, we survey the prominent electron microscopy (EM) techniques, with an emphasis on considerations for applying them to study mammalian gametes. We review how conventional EM has provided significant insight into gamete ultrastructure, but also how the harsh sample preparation methods required preclude understanding at a truly molecular level. We present recent advancements in cryo-electron tomography that provide an opportunity to image cells in a near-native state and at unprecedented levels of detail. New and emerging cellular EM techniques are poised to rekindle exploration of fundamental questions in mammalian reproduction, especially phenomena that involve complex membrane remodelling and protein reorganization. These methods will also allow novel lines of enquiry into problems of practical significance, such as investigating unexplained causes of human infertility and improving assisted reproductive technologies for biodiversity conservation.

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Section: Conventional Em Has Contributed Greatly To Our Understandingmentioning

confidence: 99%

Looking back and looking forward: contributions of electron microscopy to the structural cell biology of gametes and fertilization

2020

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“…For SEM processing of the sperm sample, spermatozoa have to undergo washings by centrifugation, fixation, dehydration through ethanols, drying by the critical point and sputter coated with a metal (Dadoune, ; Nussdorfer et al., ; Ricci et al., ). These preparing procedures may cause sperm ultrastructural modifications under certain conditions (Baccetti & Burrini, ; Chemes, ; Dong, Huang, Henk, & Tiersch, ; Grab, Thierauf, Rosenbusch, & Sterzik, ; Nussdorfer et al., ). In addition, sample observation and storing status may also affect sperm ultrastructures (Egerton, Li, & Malac, ; Jiang, ).…”

Section: Introductionmentioning

confidence: 99%

Preparation and observation methods can produce misleading artefacts in human sperm ultrastructural morphology

Zhu

2018

Andrologia

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The aim of this study was to evaluate the production of artefacts during preparation and observation of human sperm for ultrastructural morphology and analyse the possible reasons of causing these artefacts. Under the scanning electron microscopy, damaged sperm heads (crack or/and rupture), necks (head-neck or head-midpiece separation) and midpiece (disassembled and denuded axoneme ultrastructures, bent midpiece) were analysed to be the consequence of exogenous effects. Thirty infertile men with teratozoospermia revealed more spermatozoa with damage to head, neck and midpiece than did thirty fertile males (p < .01). After the samples from fertile males underwent five repeated observations, most sperm heads and necks in the samples were destroyed when compared with the single observation (p < .01). Destroyed sperm heads were full of cracks and peelings, even sperm tails were broken and fragmented, and separations of the sperm head-neck or head-midpiece became common. Spermatozoa from fertile males with centrifugation of 600 g for washing sperm exhibited more damage to the midpiece than those with the 300 g (p < .01). These results demonstrate that preparation and observation methods can damage sperm ultrastructures, leading to producing artefacts of ultrastructural morphology. The artefacts of sperm ultrastructural morphology may be associated with sperm structural fragility, preparation conditions and electron beam damage.

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Magnetic-Activated Cell Sorting of Human Spermatozoa

Dırıcan

2012

Practical Manual of in Vitro Fertilization

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Scanning electron microscopy of human sperms after preparation of semen for in-vitro fertilization

Cited by 5 publications

References 6 publications

Looking back and looking forward: contributions of electron microscopy to the structural cell biology of gametes and fertilization

Looking back and looking forward: contributions of electron microscopy to the structural cell biology of gametes and fertilization

Preparation and observation methods can produce misleading artefacts in human sperm ultrastructural morphology

Magnetic-Activated Cell Sorting of Human Spermatozoa

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