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Title of Journal: Int J Legal Med

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Abbravation: International Journal of Legal Medicine

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Springer Berlin Heidelberg

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DOI

10.1016/j.beproc.2014.07.009

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1437-1596

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Development of a quantitative validation method fo

Authors: Ayari Takamura Ken Watanabe Tomoko Akutsu
Publish Date: 2016/05/16
Volume: 130, Issue: 6, Pages: 1421-1429
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Abstract

In investigations of sexual assaults as well as in identifying a suspect the detection of human sperm is important Recently a kit for fluorescent staining of human spermatozoa SPERM HYLITER™ has become available This kit allows for microscopic observation of the heads of human sperm using an antibody tagged with a fluorescent dye This kit is specific to human sperm and provides easy detection by luminescence However criteria need to be established to objectively evaluate the fluorescent signals and to evaluate the staining efficiency of this kit These criteria will be indispensable for investigation of forensic samples In the present study the SPERM HYLITER™ Express kit which is an improved version of SPERM HYLITER™ was evaluated using an image analysis procedure using Laplacian and Gaussian methods This method could be used to automatically select important regions of fluorescence produced by sperm The fluorescence staining performance was evaluated and compared under various experimental conditions such as for aged traces and in combination with other chemical staining methods The morphological characteristics of human sperm were incorporated into the criteria for objective identification of sperm based on quantified features of the fluorescent spots Using the criteria nonspecific or insignificant fluorescent spots were excluded and the specificity of the kit for human sperm was confirmed The image analysis method and criteria established in this study are universal and could be applied under any experimental conditions These criteria will increase the reliability of operator judgment in the analysis of human sperm samples in forensics

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