Abstract

Confocal Raman microspectroscopy is a valuable analytical tool in biological and medical research, allowing the detection of sample variations without external labels or extensive preparation. To determine whether this method can assess the effect of maleic acid on sperm, we prepared human sperm samples incubated in different concentrations of maleic acid, after which Raman spectra from the various regions of sperm cells were recorded. Following the maleic acid treatment, Raman spectra indicated significant changes. Combined with other means, we found that the structures and chemical compositions of sperm membranes were damaged, and even the sperm DNA was damaged by the incorporation of maleic acid. Thus, this technique can be used for detection and identification of maleic acid-induced changes in human sperm at a molecular level. Although this particular application of Raman microspectroscopy still requires further validation, it has potentially promise as a diagnostic tool for reproductive medicine.

© 2014 Optical Society of America

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    [CrossRef] [PubMed]
  3. Z. Pandian, A. Templeton, G. Serour, and S. Bhattacharya, “Number of embryos for transfer after IVF and ICSI: a Cochrane review,” Hum. Reprod.20(10), 2681–2687 (2005).
    [CrossRef] [PubMed]
  4. D. P. Evenson, L. K. Jost, D. Marshall, M. J. Zinaman, E. Clegg, K. Purvis, P. de Angelis, and O. P. Claussen, “Utility of the sperm chromatin structure assay as a diagnostic and prognostic tool in the human fertility clinic,” Hum. Reprod.14(4), 1039–1049 (1999).
    [CrossRef] [PubMed]
  5. C. Kennedy, P. Ahlering, H. Rodriguez, S. Levy, and P. Sutovsky, “Sperm chromatin structure correlates with spontaneous abortion and multiple pregnancy rates in assisted reproduction,” Reprod. Biomed. Online22(3), 272–276 (2011).
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
  10. C. Avendaño, A. Franchi, S. Taylor, M. Morshedi, S. Bocca, and S. Oehninger, “Fragmentation of DNA in morphologically normal human spermatozoa,” Fertil. Steril.91(4), 1077–1084 (2009).
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    [CrossRef] [PubMed]
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  17. C. Matthäus, T. Chernenko, J. A. Newmark, C. M. Warner, and M. Diem, “Label-free detection of mitochondrial distribution in cells by nonresonant Raman microspectroscopy,” Biophys. J.93(2), 668–673 (2007).
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  20. M. S. Vidyasagar, K. Maheedhar, B. M. Vadhiraja, D. J. Fernendes, V. B. Kartha, and C. M. Krishna, “Prediction of radiotherapy response in cervix cancer by Raman spectroscopy: A pilot study,” Biopolymers89(6), 530–537 (2008).
    [CrossRef] [PubMed]
  21. K. C. Schuster, I. Reese, E. Urlaub, J. R. Gapes, and B. Lendl, “Multidimensional information on the chemical composition of single bacterial cells by confocal Raman microspectroscopy,” Anal. Chem.72(22), 5529–5534 (2000).
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  22. G. B. Jung, Y. J. Lee, G. Lee, and H. K. Park, “A simple and rapid detection of tissue adhesive-induced biochemical changes in cells and DNA using Raman spectroscopy,” Biomed. Opt. Express4(11), 2673–2682 (2013).
    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
  32. R. Sanchez, E. Toepfer-Petersen, R. J. Aitken, and W. B. Schill, “A new method for evaluation of the acrosome reaction in viable human spermatozoa,” Andrologia23(3), 197–203 (1991).
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
  37. J. M. Benevides and G. J. Thomas., “Characterization of DNA structures by Raman spectroscopy: high-salt and low-salt forms of double helical poly(dG-dC) in H2O and D2O solutions and application to B, Z and A-DNA,” Nucleic Acids Res.11(16), 5747–5761 (1983).
    [CrossRef] [PubMed]
  38. S. C. Erfurth and W. L. Peticolas, “Melting and premelting phenomenon in DNA by laser Raman scattering,” Biopolymers14(2), 247–264 (1975).
    [CrossRef] [PubMed]
  39. H. Deng, V. A. Bloomfield, J. M. Benevides, and G. J. Thomas., “Structural basis of polyamine-DNA recognition: spermidine and spermine interactions with genomic B-DNAs of different GC content probed by Raman spectroscopy,” Nucleic Acids Res.28(17), 3379–3385 (2000).
    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
  41. Y. Xu, Z. Zhou, H. Yang, Y. Xu, and Z. Zhang, “Raman spectroscopic study of microcosmic photodamage of the space structure of DNA sensitized by Yangzhou haematoporphyrin derivative and Photofrin II,” J. Photochem. Photobiol. B52(1-3), 30–34 (1999).
    [CrossRef] [PubMed]
  42. J. Mo, W. Zheng, and Z. Huang, “Fiber-optic Raman probe couples ball lens for depth-selected Raman measurements of epithelial tissue,” Biomed. Opt. Express1(1), 17–30 (2010).
    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
  44. B. P. Gaber and W. L. Peticolas, “On the quantitative interpretation of biomembrane structure by Raman spectroscopy,” Biochim. Biophys. Acta465(2), 260–274 (1977).
    [CrossRef] [PubMed]
  45. N. Li, S. X. Li, Z. Y. Guo, Z. F. Zhuang, R. Li, K. Xiong, S. J. Chen, and S. H. Liu, “Micro-Raman spectroscopy study of the effect of mid-ultraviolet radiation on erythrocyte membrane,” J. Photochem. Photobiol. B112, 37–42 (2012).
    [CrossRef] [PubMed]
  46. R. S. Jeyendran, H. H. Van der Ven, M. Perez-Pelaez, B. G. Crabo, and L. J. D. Zaneveld, “Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics,” J. Reprod. Fertil.70(1), 219–228 (1984).
    [CrossRef] [PubMed]
  47. D. Lechniak, A. Kedzierski, and D. Stanislawski, “The use of HOS test to evaluate membrane functionality of boar sperm capacitated in vitro,” Reprod. Domest. Anim.37(6), 379–380 (2002).
    [CrossRef] [PubMed]
  48. H. N. Sallam, A. Farrag, A.-F. Agameya, Y. El-Garem, and F. Ezzeldin, “The use of the modified hypo-osmotic swelling test for the selection of immotile testicular spermatozoa in patients treated with ICSI: a randomized controlled study,” Hum. Reprod.20(12), 3435–3440 (2005).
    [CrossRef] [PubMed]

2013 (4)

A. E. Willets, J. M. Corbo, and J. N. Brown, “Clomiphene for the treatment of male infertility,” Reprod. Sci.20(7), 739–744 (2013).
[CrossRef] [PubMed]

G. B. Jung, Y. J. Lee, G. Lee, and H. K. Park, “A simple and rapid detection of tissue adhesive-induced biochemical changes in cells and DNA using Raman spectroscopy,” Biomed. Opt. Express4(11), 2673–2682 (2013).
[CrossRef] [PubMed]

F. Liu, Y. Zhu, Y. Liu, X. Wang, P. Ping, X. Zhu, H. Hu, Z. Li, and L. He, “Real-time Raman microspectroscopy scanning of the single live sperm bound to human zona pellucida,” Fertil. Steril.99, 684–689.e684 (2013).

Z. Zhuang, N. Li, Z. Guo, M. Zhu, K. Xiong, and S. Chen, “Study of molecule variations in renal tumor based on confocal micro-Raman spectroscopy,” J. Biomed. Opt.18(3), 031103 (2013).
[CrossRef] [PubMed]

2012 (2)

N. Li, S. X. Li, Z. Y. Guo, Z. F. Zhuang, R. Li, K. Xiong, S. J. Chen, and S. H. Liu, “Micro-Raman spectroscopy study of the effect of mid-ultraviolet radiation on erythrocyte membrane,” J. Photochem. Photobiol. B112, 37–42 (2012).
[CrossRef] [PubMed]

V. Sánchez, K. Redmann, J. Wistuba, F. Wübbeling, M. Burger, H. Oldenhof, W. F. Wolkers, S. Kliesch, S. Schlatt, and C. Mallidis, “Oxidative DNA damage in human sperm can be detected by Raman microspectroscopy,” Fertil. Steril.98, 1124–1129.e1123 (2012).

2011 (3)

C. Mallidis, J. Wistuba, B. Bleisteiner, O. S. Damm, P. Gross, F. Wübbeling, C. Fallnich, M. Burger, and S. Schlatt, “In situ visualization of damaged DNA in human sperm by Raman microspectroscopy,” Hum. Reprod.26(7), 1641–1649 (2011).
[CrossRef] [PubMed]

C. Kennedy, P. Ahlering, H. Rodriguez, S. Levy, and P. Sutovsky, “Sperm chromatin structure correlates with spontaneous abortion and multiple pregnancy rates in assisted reproduction,” Reprod. Biomed. Online22(3), 272–276 (2011).
[CrossRef] [PubMed]

M. Bungum, L. Bungum, and A. Giwercman, “Sperm chromatin structure assay (SCSA): a tool in diagnosis and treatment of infertility,” Asian J. Androl.13(1), 69–75 (2011).
[CrossRef] [PubMed]

2010 (4)

K. Meister, D. A. Schmidt, E. Bründermann, and M. Havenith, “Confocal Raman microspectroscopy as an analytical tool to assess the mitochondrial status in human spermatozoa,” Analyst (Lond.)135(6), 1370–1374 (2010).
[CrossRef] [PubMed]

A. Downes, R. Mouras, and A. Elfick, “Optical spectroscopy for noninvasive monitoring of stem cell differentiation,” J. Biomed. Biotechnol.2010, 101864 (2010).
[CrossRef] [PubMed]

M. Muratori, L. Tamburrino, V. Tocci, A. Costantino, S. Marchiani, C. Giachini, I. Laface, C. Krausz, M. C. Meriggiola, G. Forti, and E. Baldi, “Small variations in crucial steps of TUNEL assay coupled to flow cytometry greatly affect measures of sperm DNA fragmentation,” J. Androl.31(4), 336–345 (2010).
[CrossRef] [PubMed]

J. Mo, W. Zheng, and Z. Huang, “Fiber-optic Raman probe couples ball lens for depth-selected Raman measurements of epithelial tissue,” Biomed. Opt. Express1(1), 17–30 (2010).
[CrossRef] [PubMed]

2009 (2)

T. Huser, C. A. Orme, C. W. Hollars, M. H. Corzett, and R. Balhorn, “Raman spectroscopy of DNA packaging in individual human sperm cells distinguishes normal from abnormal cells,” J Biophotonics2(5), 322–332 (2009).
[CrossRef] [PubMed]

C. Avendaño, A. Franchi, S. Taylor, M. Morshedi, S. Bocca, and S. Oehninger, “Fragmentation of DNA in morphologically normal human spermatozoa,” Fertil. Steril.91(4), 1077–1084 (2009).
[CrossRef] [PubMed]

2008 (1)

M. S. Vidyasagar, K. Maheedhar, B. M. Vadhiraja, D. J. Fernendes, V. B. Kartha, and C. M. Krishna, “Prediction of radiotherapy response in cervix cancer by Raman spectroscopy: A pilot study,” Biopolymers89(6), 530–537 (2008).
[CrossRef] [PubMed]

2007 (1)

C. Matthäus, T. Chernenko, J. A. Newmark, C. M. Warner, and M. Diem, “Label-free detection of mitochondrial distribution in cells by nonresonant Raman microspectroscopy,” Biophys. J.93(2), 668–673 (2007).
[CrossRef] [PubMed]

2006 (3)

P. L. Olive and J. P. Banáth, “The comet assay: a method to measure DNA damage in individual cells,” Nat. Protoc.1(1), 23–29 (2006).
[CrossRef] [PubMed]

Y. Nishimune and H. Tanaka, “Infertility caused by polymorphisms or mutations in spermatogenesis-specific genes,” J. Androl.27(3), 326–334 (2006).
[CrossRef] [PubMed]

M. V. Chowdary, K. K. Kumar, J. Kurien, S. Mathew, and C. M. Krishna, “Discrimination of normal, benign, and malignant breast tissues by Raman spectroscopy,” Biopolymers83(5), 556–569 (2006).
[CrossRef] [PubMed]

2005 (4)

Z. Pandian, A. Templeton, G. Serour, and S. Bhattacharya, “Number of embryos for transfer after IVF and ICSI: a Cochrane review,” Hum. Reprod.20(10), 2681–2687 (2005).
[CrossRef] [PubMed]

J. L. Fernández, L. Muriel, V. Goyanes, E. Segrelles, J. Gosálvez, M. Enciso, M. LaFromboise, and C. De Jonge, “Simple determination of human sperm DNA fragmentation with an improved sperm chromatin dispersion test,” Fertil. Steril.84(4), 833–842 (2005).
[CrossRef] [PubMed]

C. Krafft, T. Knetschke, R. H. W. Funk, and R. Salzer, “Identification of organelles and vesicles in single cells by Raman microspectroscopic mapping,” Vib. Spectrosc.38(1-2), 85–93 (2005).
[CrossRef]

H. N. Sallam, A. Farrag, A.-F. Agameya, Y. El-Garem, and F. Ezzeldin, “The use of the modified hypo-osmotic swelling test for the selection of immotile testicular spermatozoa in patients treated with ICSI: a randomized controlled study,” Hum. Reprod.20(12), 3435–3440 (2005).
[CrossRef] [PubMed]

2004 (3)

C. M. Krishna, G. D. Sockalingum, J. Kurien, L. Rao, L. Venteo, M. Pluot, M. Manfait, and V. B. Kartha, “Micro-Raman spectroscopy for optical pathology of oral squamous cell carcinoma,” Appl. Spectrosc.58(9), 1128–1135 (2004).
[CrossRef] [PubMed]

P. Devroey and A. Van Steirteghem, “A review of ten years experience of ICSI,” Hum. Reprod. Update10(1), 19–28 (2004).
[CrossRef] [PubMed]

V. M. Brugh and L. I. Lipshultz, “Male factor infertility,” Med. Clin. North Am.88(2), 367–385 (2004).
[CrossRef] [PubMed]

2002 (1)

D. Lechniak, A. Kedzierski, and D. Stanislawski, “The use of HOS test to evaluate membrane functionality of boar sperm capacitated in vitro,” Reprod. Domest. Anim.37(6), 379–380 (2002).
[CrossRef] [PubMed]

2001 (1)

D. S. Guzick, J. W. Overstreet, P. Factor-Litvak, C. K. Brazil, S. T. Nakajima, C. Coutifaris, S. A. Carson, P. Cisneros, M. P. Steinkampf, J. A. Hill, D. Xu, D. L. Vogel, and National Cooperative Reproductive Medicine Network, “Sperm morphology, motility, and concentration in fertile and infertile men,” N. Engl. J. Med.345(19), 1388–1393 (2001).
[CrossRef] [PubMed]

2000 (2)

K. C. Schuster, I. Reese, E. Urlaub, J. R. Gapes, and B. Lendl, “Multidimensional information on the chemical composition of single bacterial cells by confocal Raman microspectroscopy,” Anal. Chem.72(22), 5529–5534 (2000).
[CrossRef] [PubMed]

H. Deng, V. A. Bloomfield, J. M. Benevides, and G. J. Thomas., “Structural basis of polyamine-DNA recognition: spermidine and spermine interactions with genomic B-DNAs of different GC content probed by Raman spectroscopy,” Nucleic Acids Res.28(17), 3379–3385 (2000).
[CrossRef] [PubMed]

1999 (3)

H. Deng, V. A. Bloomfield, J. M. Benevides, and G. J. Thomas., “Dependence of the Raman signature of genomic B-DNA on nucleotide base sequence,” Biopolymers50(6), 656–666 (1999).
[CrossRef] [PubMed]

D. P. Evenson, L. K. Jost, D. Marshall, M. J. Zinaman, E. Clegg, K. Purvis, P. de Angelis, and O. P. Claussen, “Utility of the sperm chromatin structure assay as a diagnostic and prognostic tool in the human fertility clinic,” Hum. Reprod.14(4), 1039–1049 (1999).
[CrossRef] [PubMed]

Y. Xu, Z. Zhou, H. Yang, Y. Xu, and Z. Zhang, “Raman spectroscopic study of microcosmic photodamage of the space structure of DNA sensitized by Yangzhou haematoporphyrin derivative and Photofrin II,” J. Photochem. Photobiol. B52(1-3), 30–34 (1999).
[CrossRef] [PubMed]

1991 (1)

R. Sanchez, E. Toepfer-Petersen, R. J. Aitken, and W. B. Schill, “A new method for evaluation of the acrosome reaction in viable human spermatozoa,” Andrologia23(3), 197–203 (1991).
[CrossRef] [PubMed]

1990 (1)

G. J. Puppels, F. F. M. de Mul, C. Otto, J. Greve, M. Robert-Nicoud, D. J. Arndt-Jovin, and T. M. Jovin, “Studying single living cells and chromosomes by confocal Raman microspectroscopy,” Nature347(6290), 301–303 (1990).
[CrossRef] [PubMed]

1985 (2)

P. D. Brown-Woodman, E. J. Post, P. Y. Chow, and I. G. White, “Effects of malonic, maleic, citric and caffeic acids on the motility of human sperm and penetration of cervical mucus,” Int. J. Fertil.30(3), 38–44 (1985).
[PubMed]

A. Zaini, M. G. Jennings, and H. W. Baker, “Are conventional sperm morphology and motility assessments of predictive value in subfertile men?” Int. J. Androl.8(6), 427–435 (1985).
[CrossRef] [PubMed]

1984 (2)

H. Singh, M. S. Jabbal, A. R. Ray, and P. Vasudevan, “Effect of anionic polymeric hydrogels on spermatozoa motility,” Biomaterials5(5), 307–309 (1984).
[CrossRef] [PubMed]

R. S. Jeyendran, H. H. Van der Ven, M. Perez-Pelaez, B. G. Crabo, and L. J. D. Zaneveld, “Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics,” J. Reprod. Fertil.70(1), 219–228 (1984).
[CrossRef] [PubMed]

1983 (2)

D. M. Stein and H. Schnieden, “Effect of antidepressant drugs on the in-vitro eggpenetrating ability of golden hamster epididymal spermatozoa,” Reprod. Fertil.68(1), 227–233 (1983).
[CrossRef]

J. M. Benevides and G. J. Thomas., “Characterization of DNA structures by Raman spectroscopy: high-salt and low-salt forms of double helical poly(dG-dC) in H2O and D2O solutions and application to B, Z and A-DNA,” Nucleic Acids Res.11(16), 5747–5761 (1983).
[CrossRef] [PubMed]

1980 (1)

S. Krimm and J. Bandekar, “Vibrational analysis of peptides, polypeptides, and proteins. V. Normal vibrations of beta-turns,” Biopolymers19(1), 1–29 (1980).
[CrossRef] [PubMed]

1977 (1)

B. P. Gaber and W. L. Peticolas, “On the quantitative interpretation of biomembrane structure by Raman spectroscopy,” Biochim. Biophys. Acta465(2), 260–274 (1977).
[CrossRef] [PubMed]

1975 (1)

S. C. Erfurth and W. L. Peticolas, “Melting and premelting phenomenon in DNA by laser Raman scattering,” Biopolymers14(2), 247–264 (1975).
[CrossRef] [PubMed]

Agameya, A.-F.

H. N. Sallam, A. Farrag, A.-F. Agameya, Y. El-Garem, and F. Ezzeldin, “The use of the modified hypo-osmotic swelling test for the selection of immotile testicular spermatozoa in patients treated with ICSI: a randomized controlled study,” Hum. Reprod.20(12), 3435–3440 (2005).
[CrossRef] [PubMed]

Ahlering, P.

C. Kennedy, P. Ahlering, H. Rodriguez, S. Levy, and P. Sutovsky, “Sperm chromatin structure correlates with spontaneous abortion and multiple pregnancy rates in assisted reproduction,” Reprod. Biomed. Online22(3), 272–276 (2011).
[CrossRef] [PubMed]

Aitken, R. J.

R. Sanchez, E. Toepfer-Petersen, R. J. Aitken, and W. B. Schill, “A new method for evaluation of the acrosome reaction in viable human spermatozoa,” Andrologia23(3), 197–203 (1991).
[CrossRef] [PubMed]

Arndt-Jovin, D. J.

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K. C. Schuster, I. Reese, E. Urlaub, J. R. Gapes, and B. Lendl, “Multidimensional information on the chemical composition of single bacterial cells by confocal Raman microspectroscopy,” Anal. Chem.72(22), 5529–5534 (2000).
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R. Sanchez, E. Toepfer-Petersen, R. J. Aitken, and W. B. Schill, “A new method for evaluation of the acrosome reaction in viable human spermatozoa,” Andrologia23(3), 197–203 (1991).
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C. Mallidis, J. Wistuba, B. Bleisteiner, O. S. Damm, P. Gross, F. Wübbeling, C. Fallnich, M. Burger, and S. Schlatt, “In situ visualization of damaged DNA in human sperm by Raman microspectroscopy,” Hum. Reprod.26(7), 1641–1649 (2011).
[CrossRef] [PubMed]

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K. Meister, D. A. Schmidt, E. Bründermann, and M. Havenith, “Confocal Raman microspectroscopy as an analytical tool to assess the mitochondrial status in human spermatozoa,” Analyst (Lond.)135(6), 1370–1374 (2010).
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D. M. Stein and H. Schnieden, “Effect of antidepressant drugs on the in-vitro eggpenetrating ability of golden hamster epididymal spermatozoa,” Reprod. Fertil.68(1), 227–233 (1983).
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K. C. Schuster, I. Reese, E. Urlaub, J. R. Gapes, and B. Lendl, “Multidimensional information on the chemical composition of single bacterial cells by confocal Raman microspectroscopy,” Anal. Chem.72(22), 5529–5534 (2000).
[CrossRef] [PubMed]

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J. L. Fernández, L. Muriel, V. Goyanes, E. Segrelles, J. Gosálvez, M. Enciso, M. LaFromboise, and C. De Jonge, “Simple determination of human sperm DNA fragmentation with an improved sperm chromatin dispersion test,” Fertil. Steril.84(4), 833–842 (2005).
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Serour, G.

Z. Pandian, A. Templeton, G. Serour, and S. Bhattacharya, “Number of embryos for transfer after IVF and ICSI: a Cochrane review,” Hum. Reprod.20(10), 2681–2687 (2005).
[CrossRef] [PubMed]

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H. Singh, M. S. Jabbal, A. R. Ray, and P. Vasudevan, “Effect of anionic polymeric hydrogels on spermatozoa motility,” Biomaterials5(5), 307–309 (1984).
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Sockalingum, G. D.

Stanislawski, D.

D. Lechniak, A. Kedzierski, and D. Stanislawski, “The use of HOS test to evaluate membrane functionality of boar sperm capacitated in vitro,” Reprod. Domest. Anim.37(6), 379–380 (2002).
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D. M. Stein and H. Schnieden, “Effect of antidepressant drugs on the in-vitro eggpenetrating ability of golden hamster epididymal spermatozoa,” Reprod. Fertil.68(1), 227–233 (1983).
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D. S. Guzick, J. W. Overstreet, P. Factor-Litvak, C. K. Brazil, S. T. Nakajima, C. Coutifaris, S. A. Carson, P. Cisneros, M. P. Steinkampf, J. A. Hill, D. Xu, D. L. Vogel, and National Cooperative Reproductive Medicine Network, “Sperm morphology, motility, and concentration in fertile and infertile men,” N. Engl. J. Med.345(19), 1388–1393 (2001).
[CrossRef] [PubMed]

Sutovsky, P.

C. Kennedy, P. Ahlering, H. Rodriguez, S. Levy, and P. Sutovsky, “Sperm chromatin structure correlates with spontaneous abortion and multiple pregnancy rates in assisted reproduction,” Reprod. Biomed. Online22(3), 272–276 (2011).
[CrossRef] [PubMed]

Tamburrino, L.

M. Muratori, L. Tamburrino, V. Tocci, A. Costantino, S. Marchiani, C. Giachini, I. Laface, C. Krausz, M. C. Meriggiola, G. Forti, and E. Baldi, “Small variations in crucial steps of TUNEL assay coupled to flow cytometry greatly affect measures of sperm DNA fragmentation,” J. Androl.31(4), 336–345 (2010).
[CrossRef] [PubMed]

Tanaka, H.

Y. Nishimune and H. Tanaka, “Infertility caused by polymorphisms or mutations in spermatogenesis-specific genes,” J. Androl.27(3), 326–334 (2006).
[CrossRef] [PubMed]

Taylor, S.

C. Avendaño, A. Franchi, S. Taylor, M. Morshedi, S. Bocca, and S. Oehninger, “Fragmentation of DNA in morphologically normal human spermatozoa,” Fertil. Steril.91(4), 1077–1084 (2009).
[CrossRef] [PubMed]

Templeton, A.

Z. Pandian, A. Templeton, G. Serour, and S. Bhattacharya, “Number of embryos for transfer after IVF and ICSI: a Cochrane review,” Hum. Reprod.20(10), 2681–2687 (2005).
[CrossRef] [PubMed]

Thomas, G. J.

H. Deng, V. A. Bloomfield, J. M. Benevides, and G. J. Thomas., “Structural basis of polyamine-DNA recognition: spermidine and spermine interactions with genomic B-DNAs of different GC content probed by Raman spectroscopy,” Nucleic Acids Res.28(17), 3379–3385 (2000).
[CrossRef] [PubMed]

H. Deng, V. A. Bloomfield, J. M. Benevides, and G. J. Thomas., “Dependence of the Raman signature of genomic B-DNA on nucleotide base sequence,” Biopolymers50(6), 656–666 (1999).
[CrossRef] [PubMed]

J. M. Benevides and G. J. Thomas., “Characterization of DNA structures by Raman spectroscopy: high-salt and low-salt forms of double helical poly(dG-dC) in H2O and D2O solutions and application to B, Z and A-DNA,” Nucleic Acids Res.11(16), 5747–5761 (1983).
[CrossRef] [PubMed]

Tocci, V.

M. Muratori, L. Tamburrino, V. Tocci, A. Costantino, S. Marchiani, C. Giachini, I. Laface, C. Krausz, M. C. Meriggiola, G. Forti, and E. Baldi, “Small variations in crucial steps of TUNEL assay coupled to flow cytometry greatly affect measures of sperm DNA fragmentation,” J. Androl.31(4), 336–345 (2010).
[CrossRef] [PubMed]

Toepfer-Petersen, E.

R. Sanchez, E. Toepfer-Petersen, R. J. Aitken, and W. B. Schill, “A new method for evaluation of the acrosome reaction in viable human spermatozoa,” Andrologia23(3), 197–203 (1991).
[CrossRef] [PubMed]

Urlaub, E.

K. C. Schuster, I. Reese, E. Urlaub, J. R. Gapes, and B. Lendl, “Multidimensional information on the chemical composition of single bacterial cells by confocal Raman microspectroscopy,” Anal. Chem.72(22), 5529–5534 (2000).
[CrossRef] [PubMed]

Vadhiraja, B. M.

M. S. Vidyasagar, K. Maheedhar, B. M. Vadhiraja, D. J. Fernendes, V. B. Kartha, and C. M. Krishna, “Prediction of radiotherapy response in cervix cancer by Raman spectroscopy: A pilot study,” Biopolymers89(6), 530–537 (2008).
[CrossRef] [PubMed]

Van der Ven, H. H.

R. S. Jeyendran, H. H. Van der Ven, M. Perez-Pelaez, B. G. Crabo, and L. J. D. Zaneveld, “Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics,” J. Reprod. Fertil.70(1), 219–228 (1984).
[CrossRef] [PubMed]

Van Steirteghem, A.

P. Devroey and A. Van Steirteghem, “A review of ten years experience of ICSI,” Hum. Reprod. Update10(1), 19–28 (2004).
[CrossRef] [PubMed]

Vasudevan, P.

H. Singh, M. S. Jabbal, A. R. Ray, and P. Vasudevan, “Effect of anionic polymeric hydrogels on spermatozoa motility,” Biomaterials5(5), 307–309 (1984).
[CrossRef] [PubMed]

Venteo, L.

Vidyasagar, M. S.

M. S. Vidyasagar, K. Maheedhar, B. M. Vadhiraja, D. J. Fernendes, V. B. Kartha, and C. M. Krishna, “Prediction of radiotherapy response in cervix cancer by Raman spectroscopy: A pilot study,” Biopolymers89(6), 530–537 (2008).
[CrossRef] [PubMed]

Vogel, D. L.

D. S. Guzick, J. W. Overstreet, P. Factor-Litvak, C. K. Brazil, S. T. Nakajima, C. Coutifaris, S. A. Carson, P. Cisneros, M. P. Steinkampf, J. A. Hill, D. Xu, D. L. Vogel, and National Cooperative Reproductive Medicine Network, “Sperm morphology, motility, and concentration in fertile and infertile men,” N. Engl. J. Med.345(19), 1388–1393 (2001).
[CrossRef] [PubMed]

Wang, J.

Z. Huang, G. Chen, X. Chen, J. Wang, J. Chen, P. Lu, and R. Chen, “Rapid and label-free identification of normal spermatozoa based on image analysis and micro-Raman spectroscopy,” J Biophotonics1–5 (2013).

Wang, X.

F. Liu, Y. Zhu, Y. Liu, X. Wang, P. Ping, X. Zhu, H. Hu, Z. Li, and L. He, “Real-time Raman microspectroscopy scanning of the single live sperm bound to human zona pellucida,” Fertil. Steril.99, 684–689.e684 (2013).

Warner, C. M.

C. Matthäus, T. Chernenko, J. A. Newmark, C. M. Warner, and M. Diem, “Label-free detection of mitochondrial distribution in cells by nonresonant Raman microspectroscopy,” Biophys. J.93(2), 668–673 (2007).
[CrossRef] [PubMed]

White, I. G.

P. D. Brown-Woodman, E. J. Post, P. Y. Chow, and I. G. White, “Effects of malonic, maleic, citric and caffeic acids on the motility of human sperm and penetration of cervical mucus,” Int. J. Fertil.30(3), 38–44 (1985).
[PubMed]

Willets, A. E.

A. E. Willets, J. M. Corbo, and J. N. Brown, “Clomiphene for the treatment of male infertility,” Reprod. Sci.20(7), 739–744 (2013).
[CrossRef] [PubMed]

Wistuba, J.

V. Sánchez, K. Redmann, J. Wistuba, F. Wübbeling, M. Burger, H. Oldenhof, W. F. Wolkers, S. Kliesch, S. Schlatt, and C. Mallidis, “Oxidative DNA damage in human sperm can be detected by Raman microspectroscopy,” Fertil. Steril.98, 1124–1129.e1123 (2012).

C. Mallidis, J. Wistuba, B. Bleisteiner, O. S. Damm, P. Gross, F. Wübbeling, C. Fallnich, M. Burger, and S. Schlatt, “In situ visualization of damaged DNA in human sperm by Raman microspectroscopy,” Hum. Reprod.26(7), 1641–1649 (2011).
[CrossRef] [PubMed]

Wolkers, W. F.

V. Sánchez, K. Redmann, J. Wistuba, F. Wübbeling, M. Burger, H. Oldenhof, W. F. Wolkers, S. Kliesch, S. Schlatt, and C. Mallidis, “Oxidative DNA damage in human sperm can be detected by Raman microspectroscopy,” Fertil. Steril.98, 1124–1129.e1123 (2012).

Wübbeling, F.

V. Sánchez, K. Redmann, J. Wistuba, F. Wübbeling, M. Burger, H. Oldenhof, W. F. Wolkers, S. Kliesch, S. Schlatt, and C. Mallidis, “Oxidative DNA damage in human sperm can be detected by Raman microspectroscopy,” Fertil. Steril.98, 1124–1129.e1123 (2012).

C. Mallidis, J. Wistuba, B. Bleisteiner, O. S. Damm, P. Gross, F. Wübbeling, C. Fallnich, M. Burger, and S. Schlatt, “In situ visualization of damaged DNA in human sperm by Raman microspectroscopy,” Hum. Reprod.26(7), 1641–1649 (2011).
[CrossRef] [PubMed]

Xiong, K.

Z. Zhuang, N. Li, Z. Guo, M. Zhu, K. Xiong, and S. Chen, “Study of molecule variations in renal tumor based on confocal micro-Raman spectroscopy,” J. Biomed. Opt.18(3), 031103 (2013).
[CrossRef] [PubMed]

N. Li, S. X. Li, Z. Y. Guo, Z. F. Zhuang, R. Li, K. Xiong, S. J. Chen, and S. H. Liu, “Micro-Raman spectroscopy study of the effect of mid-ultraviolet radiation on erythrocyte membrane,” J. Photochem. Photobiol. B112, 37–42 (2012).
[CrossRef] [PubMed]

Xu, D.

D. S. Guzick, J. W. Overstreet, P. Factor-Litvak, C. K. Brazil, S. T. Nakajima, C. Coutifaris, S. A. Carson, P. Cisneros, M. P. Steinkampf, J. A. Hill, D. Xu, D. L. Vogel, and National Cooperative Reproductive Medicine Network, “Sperm morphology, motility, and concentration in fertile and infertile men,” N. Engl. J. Med.345(19), 1388–1393 (2001).
[CrossRef] [PubMed]

Xu, Y.

Y. Xu, Z. Zhou, H. Yang, Y. Xu, and Z. Zhang, “Raman spectroscopic study of microcosmic photodamage of the space structure of DNA sensitized by Yangzhou haematoporphyrin derivative and Photofrin II,” J. Photochem. Photobiol. B52(1-3), 30–34 (1999).
[CrossRef] [PubMed]

Y. Xu, Z. Zhou, H. Yang, Y. Xu, and Z. Zhang, “Raman spectroscopic study of microcosmic photodamage of the space structure of DNA sensitized by Yangzhou haematoporphyrin derivative and Photofrin II,” J. Photochem. Photobiol. B52(1-3), 30–34 (1999).
[CrossRef] [PubMed]

Yang, H.

Y. Xu, Z. Zhou, H. Yang, Y. Xu, and Z. Zhang, “Raman spectroscopic study of microcosmic photodamage of the space structure of DNA sensitized by Yangzhou haematoporphyrin derivative and Photofrin II,” J. Photochem. Photobiol. B52(1-3), 30–34 (1999).
[CrossRef] [PubMed]

Zaini, A.

A. Zaini, M. G. Jennings, and H. W. Baker, “Are conventional sperm morphology and motility assessments of predictive value in subfertile men?” Int. J. Androl.8(6), 427–435 (1985).
[CrossRef] [PubMed]

Zaneveld, L. J. D.

R. S. Jeyendran, H. H. Van der Ven, M. Perez-Pelaez, B. G. Crabo, and L. J. D. Zaneveld, “Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics,” J. Reprod. Fertil.70(1), 219–228 (1984).
[CrossRef] [PubMed]

Zhang, Z.

Y. Xu, Z. Zhou, H. Yang, Y. Xu, and Z. Zhang, “Raman spectroscopic study of microcosmic photodamage of the space structure of DNA sensitized by Yangzhou haematoporphyrin derivative and Photofrin II,” J. Photochem. Photobiol. B52(1-3), 30–34 (1999).
[CrossRef] [PubMed]

Zheng, W.

Zhou, Z.

Y. Xu, Z. Zhou, H. Yang, Y. Xu, and Z. Zhang, “Raman spectroscopic study of microcosmic photodamage of the space structure of DNA sensitized by Yangzhou haematoporphyrin derivative and Photofrin II,” J. Photochem. Photobiol. B52(1-3), 30–34 (1999).
[CrossRef] [PubMed]

Zhu, M.

Z. Zhuang, N. Li, Z. Guo, M. Zhu, K. Xiong, and S. Chen, “Study of molecule variations in renal tumor based on confocal micro-Raman spectroscopy,” J. Biomed. Opt.18(3), 031103 (2013).
[CrossRef] [PubMed]

Zhu, X.

F. Liu, Y. Zhu, Y. Liu, X. Wang, P. Ping, X. Zhu, H. Hu, Z. Li, and L. He, “Real-time Raman microspectroscopy scanning of the single live sperm bound to human zona pellucida,” Fertil. Steril.99, 684–689.e684 (2013).

Zhu, Y.

F. Liu, Y. Zhu, Y. Liu, X. Wang, P. Ping, X. Zhu, H. Hu, Z. Li, and L. He, “Real-time Raman microspectroscopy scanning of the single live sperm bound to human zona pellucida,” Fertil. Steril.99, 684–689.e684 (2013).

Zhuang, Z.

Z. Zhuang, N. Li, Z. Guo, M. Zhu, K. Xiong, and S. Chen, “Study of molecule variations in renal tumor based on confocal micro-Raman spectroscopy,” J. Biomed. Opt.18(3), 031103 (2013).
[CrossRef] [PubMed]

Zhuang, Z. F.

N. Li, S. X. Li, Z. Y. Guo, Z. F. Zhuang, R. Li, K. Xiong, S. J. Chen, and S. H. Liu, “Micro-Raman spectroscopy study of the effect of mid-ultraviolet radiation on erythrocyte membrane,” J. Photochem. Photobiol. B112, 37–42 (2012).
[CrossRef] [PubMed]

Zinaman, M. J.

D. P. Evenson, L. K. Jost, D. Marshall, M. J. Zinaman, E. Clegg, K. Purvis, P. de Angelis, and O. P. Claussen, “Utility of the sperm chromatin structure assay as a diagnostic and prognostic tool in the human fertility clinic,” Hum. Reprod.14(4), 1039–1049 (1999).
[CrossRef] [PubMed]

Anal. Chem. (1)

K. C. Schuster, I. Reese, E. Urlaub, J. R. Gapes, and B. Lendl, “Multidimensional information on the chemical composition of single bacterial cells by confocal Raman microspectroscopy,” Anal. Chem.72(22), 5529–5534 (2000).
[CrossRef] [PubMed]

Analyst (Lond.) (1)

K. Meister, D. A. Schmidt, E. Bründermann, and M. Havenith, “Confocal Raman microspectroscopy as an analytical tool to assess the mitochondrial status in human spermatozoa,” Analyst (Lond.)135(6), 1370–1374 (2010).
[CrossRef] [PubMed]

Andrologia (1)

R. Sanchez, E. Toepfer-Petersen, R. J. Aitken, and W. B. Schill, “A new method for evaluation of the acrosome reaction in viable human spermatozoa,” Andrologia23(3), 197–203 (1991).
[CrossRef] [PubMed]

Appl. Spectrosc. (1)

Asian J. Androl. (1)

M. Bungum, L. Bungum, and A. Giwercman, “Sperm chromatin structure assay (SCSA): a tool in diagnosis and treatment of infertility,” Asian J. Androl.13(1), 69–75 (2011).
[CrossRef] [PubMed]

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B. P. Gaber and W. L. Peticolas, “On the quantitative interpretation of biomembrane structure by Raman spectroscopy,” Biochim. Biophys. Acta465(2), 260–274 (1977).
[CrossRef] [PubMed]

Biomaterials (1)

H. Singh, M. S. Jabbal, A. R. Ray, and P. Vasudevan, “Effect of anionic polymeric hydrogels on spermatozoa motility,” Biomaterials5(5), 307–309 (1984).
[CrossRef] [PubMed]

Biomed. Opt. Express (2)

Biophys. J. (1)

C. Matthäus, T. Chernenko, J. A. Newmark, C. M. Warner, and M. Diem, “Label-free detection of mitochondrial distribution in cells by nonresonant Raman microspectroscopy,” Biophys. J.93(2), 668–673 (2007).
[CrossRef] [PubMed]

Biopolymers (5)

H. Deng, V. A. Bloomfield, J. M. Benevides, and G. J. Thomas., “Dependence of the Raman signature of genomic B-DNA on nucleotide base sequence,” Biopolymers50(6), 656–666 (1999).
[CrossRef] [PubMed]

S. C. Erfurth and W. L. Peticolas, “Melting and premelting phenomenon in DNA by laser Raman scattering,” Biopolymers14(2), 247–264 (1975).
[CrossRef] [PubMed]

S. Krimm and J. Bandekar, “Vibrational analysis of peptides, polypeptides, and proteins. V. Normal vibrations of beta-turns,” Biopolymers19(1), 1–29 (1980).
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M. V. Chowdary, K. K. Kumar, J. Kurien, S. Mathew, and C. M. Krishna, “Discrimination of normal, benign, and malignant breast tissues by Raman spectroscopy,” Biopolymers83(5), 556–569 (2006).
[CrossRef] [PubMed]

M. S. Vidyasagar, K. Maheedhar, B. M. Vadhiraja, D. J. Fernendes, V. B. Kartha, and C. M. Krishna, “Prediction of radiotherapy response in cervix cancer by Raman spectroscopy: A pilot study,” Biopolymers89(6), 530–537 (2008).
[CrossRef] [PubMed]

Fertil. Steril. (4)

V. Sánchez, K. Redmann, J. Wistuba, F. Wübbeling, M. Burger, H. Oldenhof, W. F. Wolkers, S. Kliesch, S. Schlatt, and C. Mallidis, “Oxidative DNA damage in human sperm can be detected by Raman microspectroscopy,” Fertil. Steril.98, 1124–1129.e1123 (2012).

F. Liu, Y. Zhu, Y. Liu, X. Wang, P. Ping, X. Zhu, H. Hu, Z. Li, and L. He, “Real-time Raman microspectroscopy scanning of the single live sperm bound to human zona pellucida,” Fertil. Steril.99, 684–689.e684 (2013).

J. L. Fernández, L. Muriel, V. Goyanes, E. Segrelles, J. Gosálvez, M. Enciso, M. LaFromboise, and C. De Jonge, “Simple determination of human sperm DNA fragmentation with an improved sperm chromatin dispersion test,” Fertil. Steril.84(4), 833–842 (2005).
[CrossRef] [PubMed]

C. Avendaño, A. Franchi, S. Taylor, M. Morshedi, S. Bocca, and S. Oehninger, “Fragmentation of DNA in morphologically normal human spermatozoa,” Fertil. Steril.91(4), 1077–1084 (2009).
[CrossRef] [PubMed]

Hum. Reprod. (4)

Z. Pandian, A. Templeton, G. Serour, and S. Bhattacharya, “Number of embryos for transfer after IVF and ICSI: a Cochrane review,” Hum. Reprod.20(10), 2681–2687 (2005).
[CrossRef] [PubMed]

D. P. Evenson, L. K. Jost, D. Marshall, M. J. Zinaman, E. Clegg, K. Purvis, P. de Angelis, and O. P. Claussen, “Utility of the sperm chromatin structure assay as a diagnostic and prognostic tool in the human fertility clinic,” Hum. Reprod.14(4), 1039–1049 (1999).
[CrossRef] [PubMed]

C. Mallidis, J. Wistuba, B. Bleisteiner, O. S. Damm, P. Gross, F. Wübbeling, C. Fallnich, M. Burger, and S. Schlatt, “In situ visualization of damaged DNA in human sperm by Raman microspectroscopy,” Hum. Reprod.26(7), 1641–1649 (2011).
[CrossRef] [PubMed]

H. N. Sallam, A. Farrag, A.-F. Agameya, Y. El-Garem, and F. Ezzeldin, “The use of the modified hypo-osmotic swelling test for the selection of immotile testicular spermatozoa in patients treated with ICSI: a randomized controlled study,” Hum. Reprod.20(12), 3435–3440 (2005).
[CrossRef] [PubMed]

Hum. Reprod. Update (1)

P. Devroey and A. Van Steirteghem, “A review of ten years experience of ICSI,” Hum. Reprod. Update10(1), 19–28 (2004).
[CrossRef] [PubMed]

Int. J. Androl. (1)

A. Zaini, M. G. Jennings, and H. W. Baker, “Are conventional sperm morphology and motility assessments of predictive value in subfertile men?” Int. J. Androl.8(6), 427–435 (1985).
[CrossRef] [PubMed]

Int. J. Fertil. (1)

P. D. Brown-Woodman, E. J. Post, P. Y. Chow, and I. G. White, “Effects of malonic, maleic, citric and caffeic acids on the motility of human sperm and penetration of cervical mucus,” Int. J. Fertil.30(3), 38–44 (1985).
[PubMed]

J Biophotonics (1)

T. Huser, C. A. Orme, C. W. Hollars, M. H. Corzett, and R. Balhorn, “Raman spectroscopy of DNA packaging in individual human sperm cells distinguishes normal from abnormal cells,” J Biophotonics2(5), 322–332 (2009).
[CrossRef] [PubMed]

J. Androl. (2)

Y. Nishimune and H. Tanaka, “Infertility caused by polymorphisms or mutations in spermatogenesis-specific genes,” J. Androl.27(3), 326–334 (2006).
[CrossRef] [PubMed]

M. Muratori, L. Tamburrino, V. Tocci, A. Costantino, S. Marchiani, C. Giachini, I. Laface, C. Krausz, M. C. Meriggiola, G. Forti, and E. Baldi, “Small variations in crucial steps of TUNEL assay coupled to flow cytometry greatly affect measures of sperm DNA fragmentation,” J. Androl.31(4), 336–345 (2010).
[CrossRef] [PubMed]

J. Biomed. Biotechnol. (1)

A. Downes, R. Mouras, and A. Elfick, “Optical spectroscopy for noninvasive monitoring of stem cell differentiation,” J. Biomed. Biotechnol.2010, 101864 (2010).
[CrossRef] [PubMed]

J. Biomed. Opt. (1)

Z. Zhuang, N. Li, Z. Guo, M. Zhu, K. Xiong, and S. Chen, “Study of molecule variations in renal tumor based on confocal micro-Raman spectroscopy,” J. Biomed. Opt.18(3), 031103 (2013).
[CrossRef] [PubMed]

J. Photochem. Photobiol. B (2)

N. Li, S. X. Li, Z. Y. Guo, Z. F. Zhuang, R. Li, K. Xiong, S. J. Chen, and S. H. Liu, “Micro-Raman spectroscopy study of the effect of mid-ultraviolet radiation on erythrocyte membrane,” J. Photochem. Photobiol. B112, 37–42 (2012).
[CrossRef] [PubMed]

Y. Xu, Z. Zhou, H. Yang, Y. Xu, and Z. Zhang, “Raman spectroscopic study of microcosmic photodamage of the space structure of DNA sensitized by Yangzhou haematoporphyrin derivative and Photofrin II,” J. Photochem. Photobiol. B52(1-3), 30–34 (1999).
[CrossRef] [PubMed]

J. Reprod. Fertil. (1)

R. S. Jeyendran, H. H. Van der Ven, M. Perez-Pelaez, B. G. Crabo, and L. J. D. Zaneveld, “Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics,” J. Reprod. Fertil.70(1), 219–228 (1984).
[CrossRef] [PubMed]

Med. Clin. North Am. (1)

V. M. Brugh and L. I. Lipshultz, “Male factor infertility,” Med. Clin. North Am.88(2), 367–385 (2004).
[CrossRef] [PubMed]

N. Engl. J. Med. (1)

D. S. Guzick, J. W. Overstreet, P. Factor-Litvak, C. K. Brazil, S. T. Nakajima, C. Coutifaris, S. A. Carson, P. Cisneros, M. P. Steinkampf, J. A. Hill, D. Xu, D. L. Vogel, and National Cooperative Reproductive Medicine Network, “Sperm morphology, motility, and concentration in fertile and infertile men,” N. Engl. J. Med.345(19), 1388–1393 (2001).
[CrossRef] [PubMed]

Nat. Protoc. (1)

P. L. Olive and J. P. Banáth, “The comet assay: a method to measure DNA damage in individual cells,” Nat. Protoc.1(1), 23–29 (2006).
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Nature (1)

G. J. Puppels, F. F. M. de Mul, C. Otto, J. Greve, M. Robert-Nicoud, D. J. Arndt-Jovin, and T. M. Jovin, “Studying single living cells and chromosomes by confocal Raman microspectroscopy,” Nature347(6290), 301–303 (1990).
[CrossRef] [PubMed]

Nucleic Acids Res. (2)

J. M. Benevides and G. J. Thomas., “Characterization of DNA structures by Raman spectroscopy: high-salt and low-salt forms of double helical poly(dG-dC) in H2O and D2O solutions and application to B, Z and A-DNA,” Nucleic Acids Res.11(16), 5747–5761 (1983).
[CrossRef] [PubMed]

H. Deng, V. A. Bloomfield, J. M. Benevides, and G. J. Thomas., “Structural basis of polyamine-DNA recognition: spermidine and spermine interactions with genomic B-DNAs of different GC content probed by Raman spectroscopy,” Nucleic Acids Res.28(17), 3379–3385 (2000).
[CrossRef] [PubMed]

Reprod. Biomed. Online (1)

C. Kennedy, P. Ahlering, H. Rodriguez, S. Levy, and P. Sutovsky, “Sperm chromatin structure correlates with spontaneous abortion and multiple pregnancy rates in assisted reproduction,” Reprod. Biomed. Online22(3), 272–276 (2011).
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Reprod. Domest. Anim. (1)

D. Lechniak, A. Kedzierski, and D. Stanislawski, “The use of HOS test to evaluate membrane functionality of boar sperm capacitated in vitro,” Reprod. Domest. Anim.37(6), 379–380 (2002).
[CrossRef] [PubMed]

Reprod. Fertil. (1)

D. M. Stein and H. Schnieden, “Effect of antidepressant drugs on the in-vitro eggpenetrating ability of golden hamster epididymal spermatozoa,” Reprod. Fertil.68(1), 227–233 (1983).
[CrossRef]

Reprod. Sci. (1)

A. E. Willets, J. M. Corbo, and J. N. Brown, “Clomiphene for the treatment of male infertility,” Reprod. Sci.20(7), 739–744 (2013).
[CrossRef] [PubMed]

Vib. Spectrosc. (1)

C. Krafft, T. Knetschke, R. H. W. Funk, and R. Salzer, “Identification of organelles and vesicles in single cells by Raman microspectroscopic mapping,” Vib. Spectrosc.38(1-2), 85–93 (2005).
[CrossRef]

Other (3)

S. Ramu and R. Jeyendran, “The Hypo-osmotic Swelling Test for Evaluation of Sperm Membrane Integrity,” in Spermatogenesis, D. T. Carrell and K. I. Aston, eds. (Humana Press, 2013).

Z. Huang, G. Chen, X. Chen, J. Wang, J. Chen, P. Lu, and R. Chen, “Rapid and label-free identification of normal spermatozoa based on image analysis and micro-Raman spectroscopy,” J Biophotonics1–5 (2013).

W. L. Peticolas, “Raman spectroscopy of DNA and proteins,” in Methods in Enzymology, S. Kenneth, ed. (Academic Press, 1995).

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Figures (8)

Fig. 1
Fig. 1

(A) Bright-field image of sperm cell and (B) averaged Raman spectra correspond to the regions of nucleus (a), middle piece (b), and acrosome (c) in normal human sperm, respectively.

Fig. 2
Fig. 2

Averaged Raman spectra obtained from the nucleus region (A), the middle piece (B), and the acrosomal region (C) of control sperm and treated sperm with different concentrations of maleic acid, respectively.

Fig. 3
Fig. 3

Spectral intensities of Raman peaks that occur in the nucleus, A: 787, 1094, 1337, 1421, 1446cm−1; in the middle piece, B: 988, 1004, 1344 cm−1; and in the acrosome, C: 1238, 1303 cm−1, after 0.01 M, 0.02 M, 0.04 M, 0.08 M and for control sperm cells without maleic acid.

Fig. 4
Fig. 4

Comparison of the spectra of control and treated sperm cells with the different concentrations of maleic acid in the region of 900 cm−1 to 1050 cm−1 from the nucleus region of human sperm..

Fig. 5
Fig. 5

(A) Comparison of the spectra of control and treated sperm cells with the different concentrations of maleic acid in the region of 800 cm−1 to 900 cm−1 from the middle piece in human sperm and (B) correlation of the maleic acid concentration with the peak intensity ratios 855/836cm−1 of the spectra.

Fig. 6
Fig. 6

PC1 vs PC2 plot for the spectra from the acrosome region in the control sperm cells (CSC) and treated sperm cells (TSC) with maleic acid incubation.

Fig. 7
Fig. 7

Evaluation of sperm membrane integrity with HOS test.

Fig. 8
Fig. 8

Flow-cytometric assessment of sperm DNA damage showing the different levels of fragmention induced by maleic acid.

Tables (1)

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Table 1 Longitudinal order-parameters in chains Strans and the rate of variation in the middle piece membrane before and after the effects of maleic acid.

Equations (1)

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strans= (I1128/I1086)sample (I1128/I1086)DPPCofsolid = (I1128/I1086)sample 1.77

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