Flow cytometry analysis of spermatozoa: Is it time for flow spermetry?

Peña, Fernando J.; Rodriguez, Jose M. Ortiz; Gil, María Concepción Robles; Ortega-Ferrusola, Cristina

doi:10.1111/rda.13261

Cited by 17 publications

(21 citation statements)

References 94 publications

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“…Flow cytometry is increasingly used in clinical andrology. Its application has been expanded to the assessment of the integrity of DNA, membrane, acrosome and mitochondrial function (Cheuquemán et al, ; Peña, Ortiz, Gil, & Ortega, ). In recent years, using different flow cytometry techniques is a more accurate way to detect different aspects of male infertility by examination of different parts of spermatozoa.…”

Section: Discussionmentioning

confidence: 99%

Effects of temperature and storage time on the motility, viability, DNA integrity and apoptosis of processed human spermatozoa

et al. 2019

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The aim of this study was to evaluate motility, viability, DNA integrity and apoptosis of spermatozoa when washed semen samples were kept for up to 12 days at 4–6°C and 25°C. In this experimental study, 26 normozoospermic semen samples were washed twice in Modified Ham's F10 and resuspended in IVF fertilisation medium. Half of the specimens were stored at 4–6°C, and the other half was kept at 25°C for 12 days. The proportions of viable, motile, spermatozoa with double‐stranded DNA and apoptotic spermatozoa were examined during storage time. Apoptosis was measured using annexin V‐PI staining followed by flow cytometry. Results showed that sperm motility and viability decreased during 12 days of sample storage (p < .001). There was no significant difference between the two temperatures in terms of motility and viability for up to 2 days (p < .05). The percentage of spermatozoa with double‐stranded DNA remained unchanged during the 12 days of storage at both temperatures (p > .05). Although there was no difference between the two temperatures in terms of motility, viability and apoptosis during the first two days of storage, storage of spermatozoa at 4–6°C is better than storage for a longer period than storage at 25°C. Sperm DNA resisted against denaturation during storage.

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Section: Discussionmentioning

confidence: 99%

Effects of temperature and storage time on the motility, viability, DNA integrity and apoptosis of processed human spermatozoa

et al. 2019

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“…The disadvantages of this probe are its low sensitivity and its quick quenching time when compared to more recently developed probes such as 5,5,6,6-tetrachloro-1,1,3,3-tetraethylbenzimidazolylcarbocyanine iodide (JC-1) or the specific MitoTracker dyes. There is a need for further investigation into the sperm mitochondrial function in the honey bee, and the use of new dyes, such as JC-1 and MitoTracker, may help in this task [113].…”

Section: Sperm Mitochondrial Function (Mitochondrial Membrane Potential)mentioning

confidence: 99%

“…To our knowledge, there are no studies evaluating the lipid peroxidation of drone sperm membranes. This analysis may be done using boron-dipyrromethene (BODIPY) probes or specific antibodies against lipid peroxides [113].…”

Section: Oxidative Stressmentioning

confidence: 99%

“…The measurement of multiple parameters simultaneously, cell by cell, may allow the development of more robust quality tests, and may expand research possibilities [113,122]. Various multiparametric tests may be developed nowadays, given the availability of a wide range of fluorescent probes, multichannel fluorescent microscopes and multi-laser cytometers.…”

Section: Multiparametric Sperm Quality Assessmentmentioning

confidence: 99%

“…Various multiparametric tests may be developed nowadays, given the availability of a wide range of fluorescent probes, multichannel fluorescent microscopes and multi-laser cytometers. In fact, numerous combinations have been described in mammals [113], and similar methods are likely to be developed in the honey bee in the near future.…”

Section: Multiparametric Sperm Quality Assessmentmentioning

confidence: 99%

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Sperm Quality Assessment in Honey Bee Drones

Yániz

Silvestre

Santolaria

2020

Biology

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The quality of honey bee drone semen is relevant in different contexts, ranging from colony productivity to pathology, toxicology and biodiversity preservation. Despite its importance, considerably less knowledge is available on this subject for the honey bee when compared to other domestic animal species. A proper assessment of sperm quality requires a multiple testing approach which discriminates between the different aspects of sperm integrity and functionality. Most studies on drone semen quality have only assessed a few parameters, such as sperm volume, sperm concentration and/or sperm plasma membrane integrity. Although more recent studies have focused on a broader variety of aspects of semen quality, some techniques currently used in vertebrates, such as computer-assisted sperm analysis (CASA) or multiparametric sperm quality testing, still remain to be developed in the honey bee. This may be attributed to the particular sperm morphology and physiology in this species, requiring the development of technologies specifically adapted to it. This article reviews the present knowledge of sperm quality in honey bee drones, highlighting its peculiarities and proposing future lines of research.

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Multiparameter Flow Cytometry Assay for Analysis of Nitrosative Stress Status in Human Spermatozoa

et al. 2020

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Male infertility is an increasing health problem, and oxidative/nitrosative stress plays an important role in the etiology of this condition. Nitrosative stress due to excessive levels of reactive nitrogen species (RNS) is associated with impaired male fertility. Flow cytometry may be a useful tool for semen evaluation, but the availability of multiparameter assays for analysis of sperm quality is limited. The present study standardized a multiparameter flow cytometry analysis for nitrosative stress status in human spermatozoa in a single assay. A suitable multicolor fluorochrome panel was designed and consisted of fluorescein‐boronate to detect peroxynitrite, a highly RNS, propidium iodide to analyze viability, tetramethylrhodamine methyl ester perchlorate to detect mitochondrial membrane potential (MMP) and monobromobimane to analyze thiol oxidation. Proper positive and negative controls for each fluorochrome were used to establish the technique, and sperm cells of different qualities and spermatozoa subjected to cryopreservation were analyzed. The results showed that the controls clearly discriminated between the high and low fluorescence intensities for each fluorochrome. The analysis of sperm cells of different quality demonstrated that the assay properly detected differences in all parameters analyzed according to sperm quality. The results may be reported as the mean fluorescence intensity of each fluorochrome and the percentage of cells exhibiting different characteristics. In conclusion, a protocol was standardized to analyze nitrosative stress status, including peroxynitrite production, viability, MMP, and thiol oxidation, in a single analysis using flow cytometry. This protocol may be applied to research approaches and clinical andrology to improve the evaluation of sperm quality and provide a promising tool to increase the use of clinical flow cytometry. © 2020 International Society for Advancement of Cytometry

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Flow cytometry analysis of spermatozoa: Is it time for flow spermetry?

Cited by 17 publications

References 94 publications

Effects of temperature and storage time on the motility, viability, DNA integrity and apoptosis of processed human spermatozoa

Effects of temperature and storage time on the motility, viability, DNA integrity and apoptosis of processed human spermatozoa

Sperm Quality Assessment in Honey Bee Drones

Multiparameter Flow Cytometry Assay for Analysis of Nitrosative Stress Status in Human Spermatozoa

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