Interferometric phase microscopy for label-free morphological evaluation of sperm cells.

OBJECTIVE To compare label-free interferometric phase microscopy (IPM) to label-free and label-based bright-field microscopy (BFM) in evaluating sperm cell morphology. This comparison helps in evaluating the potential of IPM for clinical sperm analysis without staining. DESIGN Comparison of imaging modalities. SETTING University laboratory. PATIENT(S) Sperm samples were obtained from healthy sperm donors. INTERVENTION(S) We evaluated 350 sperm cells, using portable IPM and BFM, according to World Health Organization (WHO) criteria. The parameters evaluated were length and width of the sperm head and midpiece; size and width of the acrosome; head, midpiece, and tail configuration; and general normality of the cell. MAIN OUTCOME MEASURE(S) Continuous variables were compared using the Student's t test. Categorical variables were compared with the χ(2) test of independence. Sensitivity and specificity of IPM and label-free BFM were calculated and compared with label-based BFM. RESULT(S) No statistical differences were found between IPM and label-based BFM in the WHO criteria. In contrast, IPM measurements of head and midpiece width and acrosome area were different from those of label-free BFM. Sensitivity and specificity of IPM were higher than those of label-free BFM for the WHO criteria. CONCLUSION(S) Label-free IPM can identify sperm cell abnormalities, with an excellent correlation with label-based BFM, and with higher accuracy compared with label-free BFM. Further prospective clinical trials are required to enable IPM as part of clinical sperm selection procedures.

[1]  E. Caspi,et al.  Estimating fertility potential via semen analysis data. , 1993, Human reproduction.

[2]  Nora Tischler,et al.  Experimental control of optical helicity in nanophotonics , 2014, Light: Science & Applications.

[3]  Pinhas Girshovitz,et al.  Doubling the field of view in off-axis low-coherence interferometric imaging , 2014, Light: Science & Applications.

[4]  J. Walkup,et al.  Statistical optics , 1986, IEEE Journal of Quantum Electronics.

[5]  Y. Beyth,et al.  ART success and in vivo sperm cell selection depend on the ultramorphological status of spermatozoa , 1999, Andrologia.

[6]  Simon McDowell,et al.  Advanced sperm selection techniques for assisted reproduction. , 2014, The Cochrane database of systematic reviews.

[7]  Pavel Ventruba,et al.  Digital holographic microscopy in human sperm imaging , 2011, Journal of Assisted Reproduction and Genetics.

[8]  Yona Barak,et al.  Real-time fine morphology of motile human sperm cells is associated with IVF-ICSI outcome. , 2002, Journal of andrology.

[9]  P. Devroey,et al.  Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte , 1992, The Lancet.

[10]  L. Horton,et al.  Letter: Chronic non-specific endometritis. , 1976, Lancet.

[11]  Natan T Shaked,et al.  Quantitative phase microscopy of biological samples using a portable interferometer. , 2012, Optics letters.

[12]  Gabriel Popescu,et al.  Erythrocyte structure and dynamics quantified by Hilbert phase microscopy. , 2005, Journal of biomedical optics.

[13]  S. Oehninger,et al.  Predictive value of abnormal sperm morphology in in vitro fertilization. , 1988, Fertility and sterility.

[14]  Michael W. Davidson,et al.  Differential Interference Contrast Microscopy and Modulation Contrast Microscopy , 2012 .

[15]  E. Cuche,et al.  Digital holographic microscopy: a noninvasive contrast imaging technique allowing quantitative visualization of living cells with subwavelength axial accuracy. , 2005, Optics letters.

[16]  J. Goodman Statistical Optics , 1985 .

[17]  R. Aitken,et al.  Development of a novel electrophoretic system for the isolation of human spermatozoa. , 2005, Human reproduction.

[19]  N. Raine-Fenning,et al.  Regular (ICSI) versus ultra-high magnification (IMSI) sperm selection for assisted reproduction. , 2013, The Cochrane database of systematic reviews.

[20]  F. Zernike How I discovered phase contrast. , 1955, Science.

[21]  P. C. Steptoe,et al.  BIRTH AFTER THE REIMPLANTATION OF A HUMAN EMBRYO , 1978, The Lancet.

[22]  Pinhas Girshovitz,et al.  Real-time quantitative phase reconstruction in off-axis digital holography using multiplexing. , 2014, Optics letters.

[23]  Petra Kaufmann,et al.  Two Dimensional Phase Unwrapping Theory Algorithms And Software , 2016 .

[24]  Pinhas Girshovitz,et al.  Generalized cell morphological parameters based on interferometric phase microscopy and their application to cell life cycle characterization , 2012, Biomedical optics express.

[25]  N. Shaked,et al.  Compact and portable low-coherence interferometer with off-axis geometry for quantitative phase microscopy and nanoscopy. , 2013, Optics express.

[26]  Pietro Ferraro,et al.  4D tracking of clinical seminal samples for quantitative characterization of motility parameters. , 2014, Biomedical optics express.

[27]  E. Levens,et al.  Embryo transfer practices in the United States: a survey of clinics registered with the Society for Assisted Reproductive Technology. , 2010, Fertility and sterility.