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Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
REVIEW

CASA in the medical laboratory: CASA in diagnostic andrology and assisted conception

Mathew J. Tomlinson A C and Asad Naeem B
+ Author Affiliations
- Author Affiliations

A Fertility Unit, East Block B Floor, Nottingham University Hospital, Derby Road, Nottingham, NG7 2UH, UK.

B Innovative Research Systems, 7 Newell Way, Perth, WA, 6062, Australia.

C Corresponding author. Email: mathew.tomlinson@nuh.nhs.uk

Reproduction, Fertility and Development 30(6) 850-859 https://doi.org/10.1071/RD17520
Submitted: 8 December 2017  Accepted: 15 January 2018   Published: 21 March 2018

Abstract

CASA has been used in reproductive medicine and pathology laboratories for over 25 years, yet the ‘fertility industry’ generally remains sceptical and has avoided automation, despite clear weaknesses in manual semen analysis. Early implementers had difficulty in validating CASA-Mot instruments against recommended manual methods (haemocytometer) due to the interference of seminal debris and non-sperm cells, which also affects the accuracy of grading motility. Both the inability to provide accurate sperm counts and a lack of consensus as to the value of sperm kinematic parameters appear to have continued to have a negative effect on CASA-Mot’s reputation. One positive interpretation from earlier work is that at least one or more measures of sperm velocity adds clinical value to the semen analysis, and these are clearly more objective than any manual motility analysis. Moreover, recent CASA-Mot systems offer simple solutions to earlier problems in eliminating artefacts and have been successfully validated for sperm concentration; as a result, they should be viewed with more confidence in relation to motility grading. Sperm morphology and DNA testing both require an evidence-based consensus and a well-validated (reliable, reproducible) assay to be developed before automation of either can be of real clinical benefit.

Additional keywords: morphology, motility, uncertainty.


References

Abaigar, T., Barbero, J., and Holt, W. V. (2012). Trajectory variance and autocorrelations within single-sperm tracks as population-level descriptors of sperm track complexity, predictability, and energy-generating ability. J. Androl. 33, 216–228.
Trajectory variance and autocorrelations within single-sperm tracks as population-level descriptors of sperm track complexity, predictability, and energy-generating ability.Crossref | GoogleScholarGoogle Scholar |

Aitken, R. J., Sutton, M., Warner, P., and Richardson, D. W. (1985). Relationship between the movement characteristics of human spermatozoa and their ability to penetrate cervical mucus and zona-free hamster oocytes. J. Reprod. Fertil. 73, 441–449.
Relationship between the movement characteristics of human spermatozoa and their ability to penetrate cervical mucus and zona-free hamster oocytes.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaL2M7osFCjsg%3D%3D&md5=8a4f10cc40f1a4c9056215450e8c11b8CAS |

Álvarez, C., Castilla, J. A., Ramírez, J. P., Vergara, F., Yoldi, A., Fernández, A., and Gaforio, J. J. (2005). External quality control program for semen analysis: Spanish experience. J. Assist. Reprod. Genet. 22, 379–387.
External quality control program for semen analysis: Spanish experience.Crossref | GoogleScholarGoogle Scholar |

Amann, R. P., and Katz, D. (2004). Reflections on CASA after 25 years. J. Androl. 25, 317–325.
Reflections on CASA after 25 years.Crossref | GoogleScholarGoogle Scholar |

Andersen, A. G., Ziebe, S., Jørgensen, N., Petersen, J. H., Skakkebaek, N. E., and Andersen, A. N. (2002). Time to pregnancy in relation to semen quality assessed by CASA before and after sperm separation. Hum. Reprod. 17, 173–177.
Time to pregnancy in relation to semen quality assessed by CASA before and after sperm separation.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD38%2FksFeitw%3D%3D&md5=0a9e74ebdca7aa4ac474bef352e3c1c7CAS |

Badenoch, D. F., Moore, H. D., Holt, W. V., Evans, P. R., Sidhu, B. S., and Evans, S. J. (1990). Sperm motility, velocity and migration. Br. J. Urol. 65, 204–208.
Sperm motility, velocity and migration.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK3c7pvFCgsg%3D%3D&md5=ec107518a38fd8f97db57b3dd75bbeccCAS |

Barratt, C. L. R., Tomlinson, M. J., and Cooke, I. D. (1993). Prognostic significance of computerized motility analysis for in vivo fertility. Fertil. Steril. 60, 520–525.
Prognostic significance of computerized motility analysis for in vivo fertility.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK3szoslGisQ%3D%3D&md5=36c3b9649e2bb9ea39f34fcfd3226383CAS |

Bellastella, G., Cooper, T. G., Battaglia, M., Ströse, A., Torres, I., Hellenkemper, B., Soler, C., and Sinisi, A. A. (2010). Dimensions of human ejaculated spermatozoa in Papanicolaou-stained seminal and swim-up smears obtained from the Integrated Semen Analysis System (ISAS) Asian J. Androl. 12, 871–879.
Dimensions of human ejaculated spermatozoa in Papanicolaou-stained seminal and swim-up smears obtained from the Integrated Semen Analysis System (ISAS)Crossref | GoogleScholarGoogle Scholar |

Boersma, A., Rasshofer, R., and Stolla, R. (2001). Influence of sample preparation, staining procedure and analysis conditions on bull sperm head morphometry using the morphology analyser integrated visual optical system. Reprod. Domest. Anim. 36, 222–229.
Influence of sample preparation, staining procedure and analysis conditions on bull sperm head morphometry using the morphology analyser integrated visual optical system.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD387lt1Oquw%3D%3D&md5=d7fa786a0d13ca6164a49e9842039dddCAS |

Coetzee, K., de Villiers, A., Kruger, T. F., and Lombard, C. J. (1999). Clinical value of using an automated sperm morphology analyzer (IVOS). Fertil. Steril. 71, 222–225.
Clinical value of using an automated sperm morphology analyzer (IVOS).Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK1M7ktV2itQ%3D%3D&md5=3652f354d37b0f8b1159453b5bb5b8eaCAS |

Cooper, T. G., Atkinson, A. D., and Nieschlag, E. (1999). Experience with external quality control in spermatology. Hum. Reprod. 14, 765–769.
Experience with external quality control in spermatology.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK1M3js12jsg%3D%3D&md5=5cfb6d45923c3af5a57cc78b1386bc08CAS |

Cooper, T. G., and Yeung, C. H. (2006). Computer-aided evaluation of assessment of “grade a” spermatozoa by experienced technicians. Fertil. Steril. 85, 220–224.
Computer-aided evaluation of assessment of “grade a” spermatozoa by experienced technicians.Crossref | GoogleScholarGoogle Scholar |

Cui, J., Acton, S. T., and Lin, Z. (2006). A Monte Carlo approach to rolling leukocyte tracking in vivo. Med. Image Anal. 10, 598–610.
A Monte Carlo approach to rolling leukocyte tracking in vivo.Crossref | GoogleScholarGoogle Scholar |

Davis, R. O., and Katz, D. F. (1993). Operational standards for CASA instruments. J. Androl. 14, 385–394.
| 1:STN:280:DyaK2c7hsleisg%3D%3D&md5=3d581f8d8c18a841d3c2588cc9907896CAS |

De Geyter, C., De Geyter, M., Koppers, B., and Nieschlag, E. (1998). Diagnostic accuracy of computer-assisted sperm motion analysis. Hum. Reprod. 13, 2512–2520.
Diagnostic accuracy of computer-assisted sperm motion analysis.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK1M%2FitFWisQ%3D%3D&md5=3ae8ffc8bdcf0aca3029ef627b9b768eCAS |

Farrell, P. B., Foote, R. H., and Zinaman, M. J. (1996). Motility and other characteristics of human sperm can be measured by computer-assisted sperm analysis of samples stained with Hoechst 33342. Fertil. Steril. 66, 446–453.
Motility and other characteristics of human sperm can be measured by computer-assisted sperm analysis of samples stained with Hoechst 33342.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK28zlt1KgsQ%3D%3D&md5=a1f7372ef632a2b12c5a8f00be5c9105CAS |

Freour, T., Jean, M., Mirallie, S., Langlois, M. L., Dubourdieu, S., and Barriere, P. (2009). Predictive value of CASA parameters in IUI with frozen donor sperm. Int. J. Androl. 32, 498–504.
Predictive value of CASA parameters in IUI with frozen donor sperm.Crossref | GoogleScholarGoogle Scholar |

Garrett, C., Liu, D. Y., Clarke, G. N., Rushford, D. D., and Baker, H. W. (2003). Automated semen analysis: ‘zona pellucida preferred’ sperm morphometry and straight-line velocity are related to pregnancy rate in subfertile couples. Hum. Reprod. 18, 1643–1649.
Automated semen analysis: ‘zona pellucida preferred’ sperm morphometry and straight-line velocity are related to pregnancy rate in subfertile couples.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD3szjvFagsg%3D%3D&md5=11a1625b6df0d605e36f93c81d6df526CAS |

Gefen, A. (2010). The relationship between sperm velocity and pressures applied to the zona pellucida during early sperm–oocyte penetration. J. Biomech. Eng. 132, 124501.
The relationship between sperm velocity and pressures applied to the zona pellucida during early sperm–oocyte penetration.Crossref | GoogleScholarGoogle Scholar |

Ghirelli-Filho, M., Mizrahi, F. E., Pompeo, A. C., and Glina, S. (2012). Influence of strict sperm morphology on the results of classic in vitro fertilization. Int. Braz. J. Urol. 38, 519–528.
Influence of strict sperm morphology on the results of classic in vitro fertilization.Crossref | GoogleScholarGoogle Scholar |

Hirano, Y., Shibahara, H., Obara, H., Suzuki, T., Takamizawa, S., Yamaguchi, C., Tsunoda, H., and Sato, I. (2001). Relationships between sperm motility characteristics assessed by the computer-aided sperm analysis (CASA) and fertilization rates in vitro. J. Assist. Reprod. Genet. 18, 215–220.
Relationships between sperm motility characteristics assessed by the computer-aided sperm analysis (CASA) and fertilization rates in vitro.Crossref | GoogleScholarGoogle Scholar |

Hotaling, J. M., Smith, J. F., Rosen, M., Muller, C. H., and Walsh, T. J. (2011). The relationship between isolated teratozoospermia and clinical pregnancy after in vitro fertilization with or without intracytoplasmic sperm injection: a systematic review and meta-analysis. Fertil. Steril. 95, 1141–1145.
The relationship between isolated teratozoospermia and clinical pregnancy after in vitro fertilization with or without intracytoplasmic sperm injection: a systematic review and meta-analysis.Crossref | GoogleScholarGoogle Scholar |

Ishimoto, K., and Gaffney, E. A. (2016). Mechanical tuning of mammalian sperm behaviour by hyperactivation, rheology and substrate adhesion: a numerical exploration. J. R. Soc. Interface 13, 20160633.
Mechanical tuning of mammalian sperm behaviour by hyperactivation, rheology and substrate adhesion: a numerical exploration.Crossref | GoogleScholarGoogle Scholar |

Isobe, T. (2012). New method to estimate the possibility of natural pregnancy using computer-assisted sperm analysis. Syst Biol Reprod Med 58, 339–347.
New method to estimate the possibility of natural pregnancy using computer-assisted sperm analysis.Crossref | GoogleScholarGoogle Scholar |

Jędrzejczak, P., Rzepczyńska, I., Taszarek-Hauke, G., Pawelczyk, L., Kotwicka, M., and Warchoł, W. (2005). Effect of sperm subpopulation’s kinetics on human fertilization in vitro. Arch. Androl. 51, 185–193.
Effect of sperm subpopulation’s kinetics on human fertilization in vitro.Crossref | GoogleScholarGoogle Scholar |

Keel, B. A., Quinn, P., Schmidt, C. F., Serafy, N. T., Serafy, N. T., and Schalue, T. K. (2000). Results of the American Association of Bioanalysts national proficiency testing programme in andrology. Hum. Reprod. 15, 680–686.
Results of the American Association of Bioanalysts national proficiency testing programme in andrology.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD3c7ltlGmtw%3D%3D&md5=3594117010b6617d1313ec86072e12ceCAS |

Kirkman-Brown, J. C., and Smith, D. J. (2011). Sperm motility: is viscosity fundamental to progress? Mol. Hum. Reprod. 17, 539–544.
Sperm motility: is viscosity fundamental to progress?Crossref | GoogleScholarGoogle Scholar |

Knuth, U. A., and Nieschlag, E. (1988). Comparison of computerized semen analysis with the conventional procedure in 322 patients. Fertil. Steril. 49, 881–885.
Comparison of computerized semen analysis with the conventional procedure in 322 patients.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaL1c7ps1Gjsg%3D%3D&md5=04cdef14663302a2633f69461ea08f9aCAS |

Kruger, T. F., Menkveld, R., Stander, F. S., Lombard, C. J., Van der Merwe, J. P., van Zyl, J. A., and Smith, K. (1986). Sperm morphologic features as a prognostic factor in in vitro fertilization. Fertil. Steril. 46, 1118–1123.
Sperm morphologic features as a prognostic factor in in vitro fertilization.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaL2s%2FlsVOgsg%3D%3D&md5=317e5b6cb4b7e646474576e90d0f5509CAS |

Kruger, T. F., Acosta, A. A., Simmons, K. F., Swanson, R. J., Matta, J. F., and Oehninger, S. (1988). Predictive value of abnormal sperm morphology in in vitro fertilization. Fertil. Steril. 49, 112–117.
Predictive value of abnormal sperm morphology in in vitro fertilization.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaL1c%2FotlOisQ%3D%3D&md5=1278d7b7098c949f6496ce54f4737231CAS |

Larsen, L., Scheike, T., Jensen, T. K., Bonde, J. P., Ernst, E., Hjollund, N. H., Zhou, Y., Skakkebaek, N. E., and Giwercman, A. (2000). A computer-assisted semen analysis parameters as predictors for fertility of men from the general population. The Danish First Pregnancy Planner Study Team. Hum. Reprod. 15, 1562–1567.
A computer-assisted semen analysis parameters as predictors for fertility of men from the general population. The Danish First Pregnancy Planner Study Team.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD3cvivVGlug%3D%3D&md5=f0f0ac94a26b27a2a9a3c27e590d8b92CAS |

Long, S., Woodward, B., and Tomlinson, M. J. (2017). Sperm toxicity testing: UK best practice guideline from the Association of Biomedical Andrologists. Br. J. Biomed. Sci. , .
Sperm toxicity testing: UK best practice guideline from the Association of Biomedical Andrologists.Crossref | GoogleScholarGoogle Scholar |

Malo, A. F., Gomendio, M., Garde, J., Lang-Lenton, B., Soler, A. J., and Roldan, E. R. (2006). Sperm design and sperm function. Biol. Lett. 2, 246–249.
Sperm design and sperm function.Crossref | GoogleScholarGoogle Scholar |

Morris, A. R., Coutts, J. R., and Robertson, L. (1996). A detailed study of the effect of videoframe rates of 25, 30 and 60 Hertz on human sperm movement characteristics. Hum. Reprod. 11, 304–310.
A detailed study of the effect of videoframe rates of 25, 30 and 60 Hertz on human sperm movement characteristics.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK283ls12itw%3D%3D&md5=5ebf602414d17c07d23f33747dfdf335CAS |

Mortimer, D., and Menkveld, R. (2001). Sperm morphology assessment – historical perspectives and current opinions. J. Androl. 22, 192–205.
| 1:STN:280:DC%2BD3M3lvVCitg%3D%3D&md5=aee717cc2301687b58d7e95699e9ac62CAS |

Mortimer, S. T., and Swan, M. A. (1995a). Variable kinematics of capacitating human spermatozoa. Hum. Reprod. 10, 3178–3182.
Variable kinematics of capacitating human spermatozoa.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK28vhvFKrsg%3D%3D&md5=5e1e6a72bd20524fc7ad2a494df814feCAS |

Mortimer, S. T., and Swan, M. A. (1995b). Kinematics of capacitating human spermatozoa analysed at 60 Hz. Hum. Reprod. 10, 873–879.
Kinematics of capacitating human spermatozoa analysed at 60 Hz.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK2MznvVeqtQ%3D%3D&md5=06d6a2f9502a67d3c8caea85d36b8321CAS |

Mortimer, S. T., and Swan, M. A. (1999). Effect of image sampling frequency on established and smoothing-independent kinematic values of capacitating human spermatozoa. Hum. Reprod. 14, 997–1004.
Effect of image sampling frequency on established and smoothing-independent kinematic values of capacitating human spermatozoa.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK1M3jslGqsg%3D%3D&md5=5639ca816df0c156766435296b36f029CAS |

Mortimer, D., Pandya, I. J., and Sawers, R. S. (1986). Relationship between human sperm motility characteristics and sperm penetration into human cervical mucus in vitro. J. Reprod. Fertil. 78, 93–102.
Relationship between human sperm motility characteristics and sperm penetration into human cervical mucus in vitro.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaL2s%2FhtFGmsg%3D%3D&md5=7b1a59ea00c705db7f2cf0d39e8ef7d3CAS |

Mortimer, D., Goel, N., and Shu, M. A. (1988). Evaluation of the CellSoft automated semen analysis system in a routine laboratory setting. Fertil. Steril. 50, 960–968.
Evaluation of the CellSoft automated semen analysis system in a routine laboratory setting.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaL1M%2FnvVKmtg%3D%3D&md5=a40178ab86996311dd9ffcbc4f1cccd0CAS |

Mortimer, D., Aitken, R. J., Mortimer, S. T., and Pacey, A. A. (1995). Workshop report: clinical CASA-the quest for consensus. Reprod. Fertil. Dev. 7, 951–959.
Workshop report: clinical CASA-the quest for consensus.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK283ns1alug%3D%3D&md5=8eecc54ca27aa9b17c0f7594777a3a62CAS |

Muriel, L., Meseguer, M., Fernández, J. L., Alvarez, J., Remohí, J., Pellicer, A., and Garrido, N. (2006). Value of the sperm chromatin dispersion test in predicting pregnancy outcome in intrauterine insemination: a blind prospective study. Hum. Reprod. 21, 738–744.
Value of the sperm chromatin dispersion test in predicting pregnancy outcome in intrauterine insemination: a blind prospective study.Crossref | GoogleScholarGoogle Scholar |

Oehninger, S., Franken, D. R., Sayed, E., Barroso, G., and Kolm, P. (2000). Sperm function assays and their predictive value for fertilization outcome in IVF therapy: a meta-analysis. Hum. Reprod. Update 6, 160–168.
Sperm function assays and their predictive value for fertilization outcome in IVF therapy: a meta-analysis.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD3c3kt1Klug%3D%3D&md5=bdd8235b0d9196a7d8878b38b02a2324CAS |

Punjabi, U., Wyns, C., Mahmoud, A., Vernelen, K., China, B., and Verheyen, G. (2016). Fifteen years of Belgian experience with external quality assessment of semen analysis. Andrology 4, 1084–1093.
Fifteen years of Belgian experience with external quality assessment of semen analysis.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC2s3gtFKmsg%3D%3D&md5=787951838a608e6c6d6ac0fb9c06471bCAS |

Rahman, M. S., Kwon, W. S., and Pang, M. G. (2014). Calcium influx and male fertility in the context of the sperm proteome: an update. BioMed Res. Int. 2014, Article ID 841615.
Calcium influx and male fertility in the context of the sperm proteome: an update.Crossref | GoogleScholarGoogle Scholar |

Ricci, G., Perticarari, S., Boscolo, R., Montico, M., Guaschino, S., and Presani, G. (2009). Semen preparation methods and sperm apoptosis: swim-up versus gradient-density centrifugation technique. Fertil. Steril. 91, 632–638.
Semen preparation methods and sperm apoptosis: swim-up versus gradient-density centrifugation technique.Crossref | GoogleScholarGoogle Scholar |

Rothmann, S. A., Bort, A. M., Quigley, J., and Pillow, R. (2013). Sperm morphology classification: a rational method for schemes adopted by the World Health Organization. Methods Mol. Biol. 927, 27–37.
Sperm morphology classification: a rational method for schemes adopted by the World Health Organization.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXit1Snurg%3D&md5=8d71440c7ffff9a26ccb931298df50b4CAS |

Sadeghi, S., García-Molina, A., Celma, F., Valverde, A., Fereidounfar, S., and Soler, C. (2016). Morphometric comparison by the ISAS® CASA-DNAf system of two techniques for the evaluation of DNA fragmentation in human spermatozoa. Asian J. Androl. 18, 835–839.
| 1:CAS:528:DC%2BC1cXhtlWks7Y%3D&md5=cead34056eb147fbd3e4040666e931f7CAS |

Sakkas, D., Manicardi, G. C., Tomlinson, M., Mandrioli, M., Bizzaro, D., Bianchi, P. G., and Bianchi, U. (2000). The use of two density gradient centrifugation techniques and the swim-up method to separate spermatozoa with chromatin and nuclear DNA anomalies. Hum. Reprod. 15, 1112–1116.
The use of two density gradient centrifugation techniques and the swim-up method to separate spermatozoa with chromatin and nuclear DNA anomalies.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD3c3kslejsA%3D%3D&md5=cac0cfebe03a834bdccf42e962b24921CAS |

Shibahara, H., Obara, H., Ayustawati, , Hirano, Y., Suzuki, T., Ohno, A., Takamizawa, S., and Suzuki, M. (2004). Prediction of pregnancy by intrauterine insemination using CASA estimates and strict criteria in patients with male factor infertility. Int. J. Androl. 27, 63–68.
Prediction of pregnancy by intrauterine insemination using CASA estimates and strict criteria in patients with male factor infertility.Crossref | GoogleScholarGoogle Scholar |

Sivanarayana, T., Krishna, ChR., Prakash, G. J., Krishna, K. M., Madan, K., Rani, B. S., Sudhakar, G., and Raju, G. A. (2012). CASA derived human sperm abnormalities: correlation with chromatin packing and DNA fragmentation. J. Assist. Reprod. Genet. 29, 1327–1334.
CASA derived human sperm abnormalities: correlation with chromatin packing and DNA fragmentation.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC3s7hslOiug%3D%3D&md5=e746cb6857bb33de3e76dc65efdc8d3fCAS |

Slott, V. L., Suarez, J. D., Poss, P. M., Linder, R. E., Strader, L. F., and Perreault, S. D. (1993). Optimization of the Hamilton-Thorn computerized sperm motility analysis system for use with rat spermatozoa in toxicological studies. Fundam. Appl. Toxicol. 21, 298–307.
Optimization of the Hamilton-Thorn computerized sperm motility analysis system for use with rat spermatozoa in toxicological studies.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2cXitlam&md5=d76eacfb80d9d0ce5cee3f6ff84aefa3CAS |

Soler, C., García-Molina, A., Sancho, M., Contell, J., Núñez, M., and Cooper, T. G. (2016). A new technique for analysis of human sperm morphology in unstained cells from raw semen. Reprod. Fertil. Dev. 28, 428–433.
A new technique for analysis of human sperm morphology in unstained cells from raw semen.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC28XjtFKnu7g%3D&md5=8d0b0c0e6cb481e3711d2c42c8ad6ae8CAS |

Suarez, S. S., and Pacey, A. A. (2006). Sperm transport in the female reproductive tract. Hum. Reprod. Update 12, 23–37.
Sperm transport in the female reproductive tract.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD2MnmslSgtA%3D%3D&md5=dae648d56752cdf65a71eb9589135928CAS |

Sukcharoen, N., Keith, J., Irvine, D. S., and Aitken, R. J. (1996). Prediction of the in-vitro fertilization (IVF) potential of human spermatozoa using sperm function tests: the effect of the delay between testing and IVF. Hum. Reprod. 11, 1030–1034.
Prediction of the in-vitro fertilization (IVF) potential of human spermatozoa using sperm function tests: the effect of the delay between testing and IVF.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK28zisVGrtQ%3D%3D&md5=921d08f6ee38370e163ff6cc3102056eCAS |

Tan, O., Ha, T., Carr, B. R., Nakonezny, P., Doody, K. M., and Doody, K. J. (2014). Predictive value of post-washed total progressively motile sperm count using CASA estimates in 6871 non-donor intrauterine insemination cycles. J. Assist. Reprod. Genet. 31, 1147–1153.
Predictive value of post-washed total progressively motile sperm count using CASA estimates in 6871 non-donor intrauterine insemination cycles.Crossref | GoogleScholarGoogle Scholar |

Thijssen, A., Klerkx, E., Huyser, C., Bosmans, E., Campo, R., and Ombelet, W. (2014). Influence of temperature and sperm preparation on the quality of spermatozoa. Reprod. Biomed. Online 28, 436–442.
Influence of temperature and sperm preparation on the quality of spermatozoa.Crossref | GoogleScholarGoogle Scholar |

Tomlinson, M. J. . (2016). Uncertainty of measurement and clinical value of semen analysis: has standardisation through professional guidelines helped or hindered progress? Andrology 4, 763–770.
Uncertainty of measurement and clinical value of semen analysis: has standardisation through professional guidelines helped or hindered progress?Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC2szhtlCguw%3D%3D&md5=fdce0177b1f5fdef89ae20de0d0fefe4CAS |

Tomlinson, M. J., Moffatt, O., Manicardi, G., Bizarro, D., Afnan, M., and Sakkas, D. (2001). Interrelationships between seminal parameters and sperm nuclear DNA damage before and after density gradient centrifugation: Implications for assisted conception. Hum. Reprod. 16, 2160–2165.
Interrelationships between seminal parameters and sperm nuclear DNA damage before and after density gradient centrifugation: Implications for assisted conception.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD3MrlsVerug%3D%3D&md5=bdbfaea1ebad3d21c52c0c380587fe82CAS |

Tomlinson, M. J., Pooley, K., Simpson, T., Newton, T., Hopkisson, J., Jayaprakasan, K., Jayaprakasan, R., Naeem, A., and Pridmore, T. (2010). Validation of a novel computer assisted sperm analysis (CASA) system employing multi-target tracking algorithms. Fertil. Steril. 93, 1911–1920.
Validation of a novel computer assisted sperm analysis (CASA) system employing multi-target tracking algorithms.Crossref | GoogleScholarGoogle Scholar |

Tomlinson, M. J., Lewis, S., and Morroll, D. (2013). Sperm quality and its relationship to natural and assisted conception: British Fertility Society Guidelines for Practice. Hum. Fertil. (Camb) 16, 175–193.
Sperm quality and its relationship to natural and assisted conception: British Fertility Society Guidelines for Practice.Crossref | GoogleScholarGoogle Scholar |

Tourmente, M., Gomendio, M., and Roldan, E. R. (2011). Sperm competition and the evolution of sperm design in mammals. BMC Evol. Biol. 11, 12.

van den Hoven, L., Hendriks, J. C., Verbeet, J. G., Westphal, J. R., and Wetzels, A. M. (2015). Status of sperm morphology assessment: an evaluation of methodology and clinical value. Fertil. Steril. 103, 53–58.
Status of sperm morphology assessment: an evaluation of methodology and clinical value.Crossref | GoogleScholarGoogle Scholar |

Velez de la Calle, J. F., Muller, A., Walschaerts, M., Clavere, J. L., Jimenez, C., Wittemer, C., and Thonneau, P. (2008). Sperm deoxyribonucleic acid fragmentation as assessed by the sperm chromatin dispersion test in assisted reproductive technology programs: results of a large prospective multicenter study. Fertil. Steril. 90, 1792–1799.
Sperm deoxyribonucleic acid fragmentation as assessed by the sperm chromatin dispersion test in assisted reproductive technology programs: results of a large prospective multicenter study.Crossref | GoogleScholarGoogle Scholar |

World Health Organization (WHO) (2010). WHO Laboratory Manual for the Examination and Processing of Human Semen.’ 5th edn. (WHO: Geneva.)

Yániz, J. L., Vicente-Fiel, S., Soler, C., Recreo, P., Carretero, T., Bono, A., Berné, J. M., and Santolaria, P. (2016). Comparison of different statistical approaches to evaluate morphometric sperm subpopulations in men. Asian J. Androl. 18, 819–823.

Zhu, J., Pacey, A. A., Lippes, J., and Barratt, C. L. R. (1994). The sequential effects of human cervical mucus, oviduct fluid and follicular fluid on sperm function. Fertil. Steril. 61, 1129–1135.
The sequential effects of human cervical mucus, oviduct fluid and follicular fluid on sperm function.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK2c3lt1Cgtg%3D%3D&md5=0b59f4990a4663bf8f6e4f29733288f5CAS |

Zinaman, M. J., Uhler, M. L., Vertuno, E., Fisher, S. G., and Clegg, E. D. (1996). Evaluation of computer-assisted semen analysis (CASA) with IDENT stain to determine sperm concentration. J. Androl. 17, 288–292.
| 1:STN:280:DyaK28zosVegsw%3D%3D&md5=cf96551038197d8a758ff778d69e82a3CAS |

Zini, A., Boman, J. M., Belzile, E., and Ciampi, A. (2008). Sperm DNA damage is associated with an increased risk of pregnancy loss after IVF and ICSI: systematic review and meta-analysis. Hum. Reprod. 23, 2663–2668.
Sperm DNA damage is associated with an increased risk of pregnancy loss after IVF and ICSI: systematic review and meta-analysis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXhsVWmt77I&md5=2ae2710320f2f3f467b7e82d795915e8CAS |