PURPOSE: The aim of this study was to use digital holographic microscopy (DHM) in human sperm imaging and compare quantitative phase contrast of sperm heads in normozoospermia (NZ) and oligoasthenozoospermia (OAT). METHODS: DHM spermatozoa imaging and repeated quantitative phase shift evaluation were used. Five NZ and 5 OAT samples were examined. Semen samples were examined by semen analysis and processed for DHM. Main outcome measures were maximum phase shift value of the sperm heads. Differences of the phase shift and in NZ and OAT samples were statistically tested. RESULTS: In NZ samples median phase shifts were in the range 2.72-3.21 rad and 2.00-2.15 in OAT samples. Differences among individual samples were statistically significant (p < 0.001) in both groups. Median phase shift according to sperm count was 2.90 rad in NZ samples and 2.00 rad in OAT samples. This difference was statistically significant (p < 0.001). CONCLUSION: Quantitative evaluation of the phase shift by DHM could provide new information on the exact structure and composition of the sperm head. At present, this technique is not established for clinical utility.

Department of Gynecology and Obstetrics Faculty of Medicine Masaryk University and Faculty Hospital Brno Czech Republic

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Marquet P, Rappaz B, Magistretti PJ, Cuche E, Emery Y, Colomb T, Depeursinge C. Digital holographic microscopy: a noninvasive contrast imaging technique allowing quantitative visualization of living cells with subwavelength axial accuracy. Opt Lett. 2005;30:468–470. doi: 10.1364/OL.30.000468. PubMed DOI

Vesely P, Blase C, Matouskova E, Bereiter-Hahn J. Arising podosomal structures are associated with neoplastic cell morphological phenotype induced by the microenvironment. Anticancer Res. 2006;26:967–972. PubMed

Depeursinge C, Colomb T, Emery Y, Kuhn J, Charriere F, Rappaz B, Marquet P. Digital holographic microscopy applied to life sciences. Conf Proc IEEE Eng Med Biol Soc. 2007;2007:6244–6247. PubMed

Janeckova H, Vesely P, Chmelik R. Proving tumour cells by acute nutritional/energy deprivation as a survival threat: a task for microscopy. Anticancer Res. 2009;29:2339–2345. PubMed

Kolman P, Chmelik R. Coherence-controlled holographic microscope. Opt Express. 2010;18:21990–22003. doi: 10.1364/OE.18.021990. PubMed DOI

Chmelik R, Harna Z. Parallel-mode confocal microscope. Opt Eng. 1999;38:1635–1639. doi: 10.1117/1.602217. DOI

Chmelik R. Three-dimensional scalar imaging in high-aperture low-coherence interference and holographic microscopes. J Mod Opt. 2006;53:2673–2689. doi: 10.1080/09500340600828541. DOI

Dubois F, Joannes L, Legros JC. Improved three-dimensional imaging with a digital holography microscope with a source of partial spatial coherence. Appl Opt. 1999;38:7085–7094. doi: 10.1364/AO.38.007085. PubMed DOI

Janeckova H, Kolman P, Vesely P, Chmelik R. Digital holographic microscope with low-spatial and temporal coherence of illumination. In: Optical and Digital Image Processing. Proc. of SPIE 2008; 7000:1–8.

World Health Organization Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interaction. 4th ed, 1999. Cambridge University Press.

Crha I, Kralikova M, Melounova J, Ventruba P, Zakova J, Beharka R, Husicka R, Pohanka M, Huser M. Seminal plasma homocysteine, folate and cobalamin in men with obstructive and non-obstructive azoospermia. J Assist Reprod Genet. 2010;27:533–538. doi: 10.1007/s10815-010-9458-8. PubMed DOI PMC

Oliva R. Protamines and male infertility. Hum Reprod Update. 2006;12:417–435. doi: 10.1093/humupd/dml009. PubMed DOI

Lefièvre L, Bedu-Addo K, Conner SJ, Machado-Oliveira GS, Chen Y, Kirkman-Brown JC, Afnan MA, Publicover SJ, Ford WC, Barratt CL. Counting sperm does not add up any more: time for a new equation? Reproduction. 2007;133:675–684. doi: 10.1530/REP-06-0332. PubMed DOI

Zini A, Phillips S, Courchesne A, Boman JM, Baazeem A, Bissonnette F, Kadoch IJ, San Gabriel M. Sperm head morphology is related to high deoxyribonucleic acid stainability assessed by sperm chromatin structure assay. Fertil Steril. 2009;91:2495–2500. doi: 10.1016/j.fertnstert.2008.03.032. PubMed DOI

Mudrak O, Tomilin N, Zalensky A. Chromosome architecture in the decondensing human sperm nucleus. J Cell Sci. 2005;118:4541–4550. doi: 10.1242/jcs.02581. PubMed DOI PMC

Ward WS. Function of sperm chromatin structural elements in fertilization and development. Mol Hum Reprod. 2010;16:30–36. doi: 10.1093/molehr/gap080. PubMed DOI PMC

Moskovtsev SI, Willis J, Azad A, Mullen JB. Sperm DNA integrity: correlation with sperm plasma membrane integrity in semen evaluated for male infertility. Arch Androl. 2005;51:33–40. doi: 10.1080/014850190512770. PubMed DOI

Akgul M, Ozkinay F, Ercal D, Cogulu O, Dogan O, Altay B, Tavmergen E, Gunduz C, Ozkinay C. Cytogenetic abnormalities in 179 cases with male infertility in Western Region of Turkey: Report and review. J Assist Reprod Genet. 2009;26:119–122. doi: 10.1007/s10815-009-9296-8. PubMed DOI PMC

Winkle T, Rosenbusch B, Gagsteiger F, Paiss T, Zoller N. The correlation between male age, sperm quality and sperm DNA fragmentation in 320 men attending a fertility center. J Assist Reprod Genet. 2009;26:41–46. doi: 10.1007/s10815-008-9277-3. PubMed DOI PMC

García-Herrero S, Garrido N, Martínez-Conejero JA, Remohí J, Pellicer A, Meseguer M. Ontological evaluation of transcriptional differences between sperm of infertile males and fertile donors using microarray analysis. J Assist Reprod Genet. 2010;27:111–120. doi: 10.1007/s10815-010-9388-5. PubMed DOI PMC

Gianaroli L, Magli MC, Ferraretti AP, Crippa A, Lappi M, Capitani S, Baccetti B. Birefringence characteristics in sperm heads allow for the selection of reacted spermatozoa for intracytoplasmic sperm injection. Fertil Steril. 2010;93:807–813. doi: 10.1016/j.fertnstert.2008.10.024. PubMed DOI

Petersen CG, Vagnini LD, Mauri AL, Massaro FC, Cavagna M, Baruffi RL, Oliveira JB, Franco JG Jr. Relationship between DNA damage and sperm head birefringence. Reprod Biomed Online. 2011 Mar 21. [Epub ahead of print] doi:10.1016/j.rbmo.2011.03.017 PubMed

Hammadeh ME, Bernardi A, Zeginiadou T, Amer A, Schmidt W. Relationship between nuclear chromatin decondensation (NCD) in vitro and other sperm parameters and their predictive value on fertilization rate in IVF program. J Assist Reprod Genet. 2005;22:301–305. doi: 10.1007/s10815-005-6002-3. PubMed DOI

Kolle S, Reese S, Kummer W. New aspects of gamete transport, fertilization, and embryonic development in the oviduct gained by means of live cell imaging. Theriogenology. 2010;73:786–795. doi: 10.1016/j.theriogenology.2009.11.002. PubMed DOI

Ierardi V, Niccolini A, Alderighi M, Gazzano A, Martelli F, Solaro R. AFM characterization of rabbit spermatozoa. Microsc Res Tech. 2008;71:529–535. doi: 10.1002/jemt.20584. PubMed DOI