Fluorescence in situ hybridization (FISH) protocol in human sperm
Anton, Ester (Universitat Autònoma de Barcelona. Departament de Biologia Cel·lular, de Fisiologia i d'Immunologia)
Sarrate Navas, Zaida (Universitat Autònoma de Barcelona. Departament de Biologia Cel·lular, de Fisiologia i d'Immunologia)

Data:	2009
Resum:	Aneuploidies are the most frequent chromosomal abnormalities in humans. Most of these abnormalities result from meiotic errors during the gametogenic process in the parents. In human males, these errors can lead to the production of spermatozoa with numerical chromosome abnormalities which represent an increased risk of transmitting these anomalies to the offspring. For this reason, the technique of fluorescence in situ hybridization (FISH) on sperm nuclei has become a protocol widely incorporated in the context of clinical diagnosis. This practice provides an estimate of the frequencies of numerical chromosome abnormalities in the gametes of the patients that seek for genetic reproductive advice. To date, the chromosomes most frequently included in sperm FISH analysis are chromosomes X, Y, 13, 18 and 21. This video-article describes, step by step, how to process and fix a human semen sample, how to decondense and denature the sperm chromatin, how to proceed to obtain sperm FISH preparations, and how to visualize the results at the microscope. Special remarks of the most relevant steps are given to achieve the best results.
Ajuts:	Agència de Gestió d'Ajuts Universitaris i de Recerca 2005/SGR-00437
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Llengua:	Anglès
Document:	Article ; recerca ; Versió acceptada per publicar
Matèria:	Cellular Biology ; Fluorescence in situ hybridization ; Human ; Infertility ; Issue 31 ; Numerical chromosome abnormalities ; Spermatozoa
Publicat a:	Journal of visualized experiments, Núm. 31 (2009) , art. e1405, ISSN 1940-087X

DOI: 10.3791/1405
PMID: 19724245

11 p, 406.8 KB

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