Raman micro-spectroscopy analysis of different sperm regions: a species comparison

Amaral, Sandra; Costa, Raul Da; Wübbeling, Frank; Redmαnn, K.; Schlatt, Stefan

doi:10.1093/molehr/gax071

Cited by 16 publications

(11 citation statements)

References 47 publications

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“…The mean spectrum of each treatment was obtained and peak assignment was done having as reference the works of Talari et al [ 29 ] and Amaral et al . [ 26 ] which we highly recommend for further details about the spectral profile of sperm cells. The measurements of all individual spectra (over the 650–1,800 cm-1 spectral range) were analyzed by Standard principal component analysis (PCA) and plotted using the R 3.4.1 and RStudio 1.0.153 [ 30 ].…”

Section: Methodsmentioning

confidence: 99%

“…This technique has also been applied for the non-invasive sex assessment in bovine semen, being able to distinguish X and Y bearing sperm [ 24 , 25 ]. Recently our group reported that several organelle-specific (sperm head, midpiece and tail) spectral features are present in human, monkey, mouse and sea urchin sperm, which could be considered potential spectral markers for sperm quality [ 26 ].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Spectral features of nuclear DNA in human sperm assessed by Raman Microspectroscopy: Effects of UV-irradiation and hydration

Costa¹,

Amaral²,

Redmαnn³

et al. 2018

PLoS ONE

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Raman Microspectroscopy represents an innovative tool for the assessment of sperm biochemical features otherwise undetectable by routine semen analysis. Previously, it was shown that induced DNA damage can be detected in smeared sperm by this technique. This novel readout may be of value for clinical settings especially if it can be transferred to living cells. Yet, starting with living sperms this study was carried-out using a variety of conditions to disclose the Raman features of sperm nuclei under different hydration conditions and UV exposure. Human sperm were immobilized and Raman spectra were obtained from individual sperm as repeated measurements. To create conditions with controlled DNA damage, sperm samples were exposed to ultraviolet light. Several media were used to evaluate their effect on Raman spectra in aqueous conditions. To substantiate differences between the experimental conditions, the spectra were analyzed by Principal Component Analysis. We observed that spectra of sperm nuclei obtained in different solutions showed a qualitatively unchanged spectral pattern showing the principal signals related to DNA. Evaluating the effect of ultraviolet light generated the finding that spectra representing DNA damage were only observed in dry conditions but not in aqueous medium. Thus, Raman microspectroscopy was successfully applied for sperm analysis in different conditions, among them in live spermatozoa in aqueous solution during the initial measurement, revealing the principle use of this technique. However, implementation of Raman spectroscopy as a technique for clinical sperm analysis and selection may be especially relevant when DNA evaluation can be established using live sperm.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Spectral features of nuclear DNA in human sperm assessed by Raman Microspectroscopy: Effects of UV-irradiation and hydration

Costa¹,

Amaral²,

Redmαnn³

et al. 2018

PLoS ONE

Self Cite

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show abstract

“…Research has so far presented controversial data on sperm evaluation, and except for nuclear DNA, the identification and assignment of spectral bands in Raman-profiles to the different sperm regions remains to be established ( Amaral et al ., 2018 ). Raman microspectroscopy is a promising tool in the assessment of male fertility, but studies are, unfortunately, in their infancy, which may delay its potential use in IVF.…”

Section: Resultsmentioning

confidence: 99%

Sperm selection in IVF: the long and winding road from bench to bedside

Pedrosa¹,

Furtado

Ferreira³

et al. 2020

JBRA Assisted Reproduction

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Spermatozoa wage battle to conquer fertilization but the traits needed to succeed remain elusive. The natural advantageous qualities that enable only a few selected sperm cells to reach the site of fertilization remain unknown. Although in vitro fertilization (IVF) facilitates the job of spermatozoa, a universally acceptable means of sperm selection is yet to be developed. No objective or reliable sperm quality indicators have been established and sperm selection is, to a great extent, based on subjective qualitative evaluation. The best method for sperm selection in IVF presents several challenges: intrinsic sperm qualities cannot be evaluated and the ideal endpoint for these studies is debatable. An ideal method for sperm selection in ART should be noninvasive and cost-effective, and allow the identification of high-quality spermatozoa and yield better outcomes in terms of pregnancy and live birth rates. This narrative review included 85 papers and focused on the new available methods and technologies that might shed some light on sperm selection in IVF. It discusses the available data on microfluidic devices, omics profiling, micronuclei studies, sperm plasma membrane markers, and other techniques, such as Magnetic Activated Cell Sorting (MACS), Raman micro-spectroscopy, and artificial intelligence systems. The new techniques herein reviewed offer fresh approaches to an old problem, for which a definite solution has yet to cross the bridge from bench to IVF clinics around the world, since clinical usefulness and application remain unproven.

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“…To establish a new diagnostic technique based on the Raman spectroscopy that is able to distinguish between sperm with normal and abnormal chromatin condensation it is essential to understand the Raman spectra of the sperm components and Raman spectral differences between normal sperm and sperm with chromatin condensation abnormalities. Previous studies analyzed Raman spectra and correlated spectral features to various properties [25,26,[39][40][41][42][45][46][47][48][49][50][51][52]. The presence of the Raman peak around 670 cm -1 represents the ring breathing mode of guanine, indicates that the DNA backbone was in B-form conformation [53].…”

Section: Discussionmentioning

confidence: 99%

Label-Free Evaluation of Chromatin Condensation in Human Normal Morphology Sperm Using Raman Spectroscopy

et al. 2021

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Chromatin condensation is one of the main factors essential for sperm function. Evaluation of chromatin condensation by current methods render the assessed sperm unsuitable for assisted reproduction. We examined the Raman spectra of normal morphology sperm to determine whether a non-invasive confocal Raman spectroscopy can detect spectral differences between groups having different levels of chromatin condensation. Semen samples from 85 donors who underwent ICSI were obtained. Chromomycin A3, aniline blue and acridine orange staining were performed to evaluate the protamine deficiency, histone retention and DNA fragmentation respectively. Raman spectra were obtained from 50 normal morphology sperm for each donor. Spectral analysis was performed using home written programs in LabVIEW software and samples were grouped based on chromomycin A3 staining. Raman peaks intensities at 670 cm-1, 731 cm-1, 785 cm-1, 858 cm-1, 1062 cm-1, 1098 cm-1, 1185 cm-1, 1372 cm-1, 1424 cm-1, 1450 cm-1, 1532 cm-1, 1618 cm-1 and 1673 cm-1 were significantly correlated with at least one of the sperm staining methods. The median intensity of the Raman peaks at 670 cm-1, 731 cm-1, 785 cm-1, 1062 cm-1, 1098 cm-1, 1185 cm-1, 1372 cm-1, 1424 cm-1, 1450 cm-1, 1532 cm-1, 1618 cm-1 and 1673 cm-1 show a significant difference between the CMA3≤41 and CMA3>41groups. The Raman spectroscopic measurements represent a promising diagnostic tool that has the ability to label-free detect sperm with chromatin abnormalities, such as improper chromatin condensation and DNA fragmentation to a certain degree similar to that of the existing staining techniques at the individual cell level.

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Raman micro-spectroscopy analysis of different sperm regions: a species comparison

Cited by 16 publications

References 47 publications

Spectral features of nuclear DNA in human sperm assessed by Raman Microspectroscopy: Effects of UV-irradiation and hydration

Spectral features of nuclear DNA in human sperm assessed by Raman Microspectroscopy: Effects of UV-irradiation and hydration

Sperm selection in IVF: the long and winding road from bench to bedside

Label-Free Evaluation of Chromatin Condensation in Human Normal Morphology Sperm Using Raman Spectroscopy

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