الفهرس 
. IntroductionOnly 14 pages are availabe for public view
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Abstract
Assisted reproductive technologies (ART) revolutionized human fertility by providing treatment to patients previously considered incapable for reproductive purposes. Intracytoplasmic sperm injection (ICSI), while typically effective for overcoming low or absent fertilization in couples with an apparent abnormality of semen parameters, is the most frequently used ART worldwide. ICSI is being used commonly in many cases such as: unexplained infertility, poor sperm quality, low sperm count, poor oocytes quality and low oocyte yield. Also, advanced maternal age, repeated failure of pregnancy, preimplantation genetic testing, fertilization after in vitro maturation, and fertilization of cryopreserved oocytes. Preselection of the gender of offspring is a subject that has held man’s attention since the beginning of recorded history. Three hypotheses for preselection of the gender are based on physical characteristics of sperm, one on supposed differences in size and shape, another on differences in density, and a third on differences in surface charge. Sorting of X (female) or Y (male) chromosome-bearing spermatozoa in semen is a popular method for sex-selection. A variety of separation methods, including magnetic separation. Several techniques are used to prepare the sperm for ART. Density gradient centrifugation is the most common method used to process semen in assisted reproduction techniques. Sperm processing by the discontinuous concentration gradient technique usually recovers a clean fraction of highly motile spermatozoa, thus separating spermatozoa from ejaculates with very low sperm density and providing good yield The present study collected 60 semen samples from men where their semen samples were analyzed; samples were obtained by masturbating, and after liquefaction, they were analyzed using an intelligent CASA program and a phase-contrast microscope. Samples were prepared using density gradient centrifugation and magnetic-activated cell sorting and measured using fluorescent in situ hybridization. Density gradient centrifugation separates sperm cells based on their density. Thus, at the end of centrifugation, each spermatozoon is located at the gradient level that matches its density. Magnetic-activated cell sorting (MACS) is an excellent tool for separating cells of interest out of mixed cell populations. The sorter utilizes magnetic micro-/nanoparticles conjugated with antibodies specific to the cell membrane protein of interest. A sperm cell with a Y chromosome is distinguishable from an X-carrying sperm by its bright fluorescent spot. Only flow cytometry (FCM) has proven effective based on the 2.8% total DNA content difference between X- and Y-bearing spermatozoa. Fluorescence in-situ hybridization (FISH) has been the most effective method for evaluating sorted spermatozoa and studying aneuploidy. This study’s authors investigated FISH’s efficacy using probes specific to X and Y chromosomes. These probes were directly labeled with different color fluorophores so that the X and Y chromosome signals could be observed simultaneously. Also, the study evaluated two techniques that were measured by the FISH technique in comparison to the standard in sex-sorting of the sperm for assessment of the efficacy of each one of them. In the current study showed that the percent of X- bearing sperms obtained by wash technique was 53.8 %, by DGC technique was 62.6 % and by MACS technique was 50.9 % respectively. Present results showed that the percent of Y- bearing sperms obtained by wash technique was 46.3 %, by DGC technique was 37.4 % and by MACS technique was 49.2 % respectively. This is the first study to compare DGC and MACS technique in sex- sorting of the sperms and to assess the agreement of the results of different methods. The value of MACS in sperm selection and its effect on the quality of the outcome had been evaluated before, so the results of this study showed that the MACS method, by separating the sperms is an excellent and noninvasive method to determine gender in fertile couples. By employing the above process, the number of unwanted pregnancies or abortions decreases, and diseases related to sex can be prevented.”