The staining pattern of human sperm with Diff Quik: relationship with sperm head morphology and a sperm chromatin structure assay (SCSA)

Moska, Natalie; Murray, Elaine; Wakefield, Penelope; Matson, Phillip

doi:10.1016/s1642-431x(12)60064-3

Cited by 3 publications

(4 citation statements)

References 11 publications

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“…Sperm Morphology Analysis : Sperm were collected from the outlet and evaluated morphologically using Quick III sperm morphology kit 74. Briefly, sperm were collected at the channel outlet 30 min after loading into the inlet, and smeared on the glass slide.…”

Section: Methodsmentioning

confidence: 99%

Guidance and Self‐Sorting of Active Swimmers: 3D Periodic Arrays Increase Persistence Length of Human Sperm Selecting for the Fittest

et al. 2017

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Male infertility is a reproductive disease, and existing clinical solutions for this condition often involve long and cumbersome sperm sorting methods, including preprocessing and centrifugation‐based steps. These methods also fall short when sorting for sperm free of reactive oxygen species, DNA damage, and epigenetic aberrations. Although several microfluidic platforms exist, they suffer from structural complexities, i.e., pumps or chemoattractants, setting insurmountable barriers to clinical adoption. Inspired by the natural filter‐like capabilities of the female reproductive tract for sperm selection, a model‐driven design, featuring pillar arrays that efficiently and noninvasively isolate highly motile and morphologically normal sperm, with lower epigenetic global methylation, from raw semen, is presented. The Simple Periodic ARray for Trapping And isolatioN (SPARTAN) created here modulates the directional persistence of sperm, increasing the spatial separation between progressive and nonprogressive motile sperm populations within an unprecedentedly short 10 min assay time. With over 99% motility of sorted sperm, a 5‐fold improvement in morphology, 3‐fold increase in nuclear maturity, and 2–4‐fold enhancement in DNA integrity, SPARTAN offers to standardize sperm selection while eliminating operator‐to‐operator variations, centrifugation, and flow. SPARTAN can also be applied in other areas, including conservation ecology, breeding of farm animals, and design of flagellar microrobots for diagnostics.

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Section: Methodsmentioning

confidence: 99%

Guidance and Self‐Sorting of Active Swimmers: 3D Periodic Arrays Increase Persistence Length of Human Sperm Selecting for the Fittest

et al. 2017

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“…Conversely, Moska et al . () concluded that the dark staining seen with Diff‐Quik is not associated with changes in human sperm chromatin detected by the SCSA. Diff‐Quik staining is therefore not a useful alternative to the SCSA.…”

Section: Discussionmentioning

confidence: 97%

“…DQ presents particular advantages as it is routinely employed in clinics for bacteriological and cellular purposes, it is a rapid, simple and inexpensive procedure, and it can be used for concurrent assessment of sperm morphology (Sousa et al ., ). Therefore, the assessment of both sperm morphology and chromatin on the same slide would be attractive to andrology laboratories (Moska et al ., ).…”

Section: Discussionmentioning

confidence: 97%

Freeze-dried stallion spermatozoa: evaluation of two chelating agents and comparative analysis of three sperm DNA damage assays

et al. 2016

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During the freeze-drying procedure, sperm DNA might become damaged by both freezing and drying stresses. Sperm DNA status can be detected using well-established assays; however, most techniques are expensive and involve elaborate protocols and equipment. Indirect assessments can provide alternative strategies. The objective of this study was to compare a simple test of DNA status using Diff-Quik (DQ) with two established procedures: acridine orange test (AOT) and sperm chromatin dispersion (SCD) on freeze-dried (FD) stallion spermatozoa. Ejaculated spermatozoa from three stallions were freeze-dried in basic medium supplemented with two different chelating agents: EGTA or EDTA. After rehydration, the spermatozoa were subjected to DNA damage detection using a SCDt, AOT and DQ stain simultaneously. The results showed that the DNA damage levels in the EGTA group were significantly lower than those in the EDTA group. AOT detected a significantly higher proportion of spermatozoa with fragmented DNA than DQ and SCD. The results of the SCD test and DQ stain exhibited a significant positive correlation for DNA fragmentation (r = 0.528), whereas a negative correlation was observed between SCD, DQ and AOT (r = -0.134 and r = -0.332 respectively). The present study shows that both the SCD test and DQ assay are effective methods for detecting FD stallion sperm DNA fragmentation, whereas using of AOT is questionable.

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“…Briefly, a 5-μl drop of the sperm sample was smeared, air-dried and stained with the kit. More than 100 spermatozoa were observed under the microscope with bright field illumination according to the criteria of morphological normality of sperm head (Moska et al , 2011). In fresh ejaculates from the patients (Table S1), the volume of ejaculate and sperm concentration were 2.0–4.1 ml (average 2.9 ml) and 46.0–218.0 × 10 6 cells/ml (average 128.1 × 10 6 cells/ml), respectively.…”

Section: Methodsmentioning

confidence: 99%

Individual differences in the distribution of sperm acrosome-associated 1 proteins among male patients of infertile couples; their possible impact on outcomes of conventional in vitro fertilization

Kishida

Harayama

Kimura

et al. 2016

Zygote

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The aims of this study were to show the existence of individual differences in the distribution of sperm acrosome-associated 1 (SPACA1) among male patients of infertile couples and to examine their possible impact on the outcomes of conventional in vitro fertilization (IVF). The spermatozoa were collected from male patients of infertile couples, washed by centrifugation, collected by the swim-up method, and then used for clinical treatments of conventional IVF. The surplus sperm samples were fixed and stained with an anti-SPACA1 polyclonal antibody for the immunocytochemistry. In the clinical IVF treatments, fertilization rates and blastocyst development rates were evaluated. The immunocytochemical observations revealed that SPACA1 were localized definitely in the acrosomal equatorial segment and variedly in the acrosomal principal segment. Specifically, the detection patterns of SPACA1 in the acrosomal principal segment could be classified into three categories: (A) strong, (B) intermediate or faint, and (C) almost no immunofluorescence. The SPACA1 indexes were largely different among male patients with the wide range from 13 to 199 points. The SPACA1 indexes were significantly correlated with developmental rates of embryos to blastocysts (r = 0.829, P = 0.00162), although they were barely associated with fertilization rates at 19 h after insemination (r = 0.289, P = 0.389). These results suggest that the distribution of SPACA1 in sperm affects the outcomes of conventional IVF. In conclusion, this study provides initial data to promote large-scale clinical investigation to demonstrate that the SPACA1 indexes are valid as molecular biomarkers that can predict the effectiveness of conventional IVF of infertile couples.

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The staining pattern of human sperm with Diff Quik: relationship with sperm head morphology and a sperm chromatin structure assay (SCSA)

Cited by 3 publications

References 11 publications

Guidance and Self‐Sorting of Active Swimmers: 3D Periodic Arrays Increase Persistence Length of Human Sperm Selecting for the Fittest

Guidance and Self‐Sorting of Active Swimmers: 3D Periodic Arrays Increase Persistence Length of Human Sperm Selecting for the Fittest

Freeze-dried stallion spermatozoa: evaluation of two chelating agents and comparative analysis of three sperm DNA damage assays

Individual differences in the distribution of sperm acrosome-associated 1 proteins among male patients of infertile couples; their possible impact on outcomes of conventional in vitro fertilization

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