Optimization of spermatozoa detection using immunofluorescent staining and laser micro-dissection

Forensic Science International : Synergy
Yueh Shyang PingChristopher Kiu Choong Syn

Abstract

The present study evaluated the use of an immunofluorescence-based assay for the microscopic detection of human spermatozoa, following which the fluorescence-labelled spermatozoa could be excised with a laser micro-dissection system. The Sperm Hy-Liter™ PI kit was able to detect spermatozoa from as little as 20nL of semen. No interference or non-specificity were observed when the kit was used on semen mixed with various body fluids such as blood and urine, as well as when semen was spiked onto different types of fabric. Good results could also be obtained with rectal samples which contain auto-fluorescent fecal materials through the use of dual FITC/PI filters. We also developed a method for concurrent testing of two protein biomarkers of semen (semenogelin and prostate-specific antigen) and detection of spermatozoa. This approach would maximize the evidential value from a single piece of sexual assault exhibit. The results also showed that staining by Sperm Hy-Liter™ PI does not interfere with DNA recovery, facilitating the generation of clear male DNA profiles from dissected spermatozoa, thereby making profile interpretation less complex. In summary, Sperm Hy-Liter™ PI staining was demonstrated to be sensitive, robust and spe...Continue Reading

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Citations

Feb 7, 2012·Forensic Science International : Synergy·Danielle GrayBarbara Daniel
Apr 11, 2013·Forensic Science International. Genetics·Anick De MoorsChantal J Frégeau
May 16, 2012·Science & Justice : Journal of the Forensic Science Society·J LewisR Talbot
May 16, 2012·Science & Justice : Journal of the Forensic Science Society·G Davidson, T B Jalowiecki
Nov 7, 2019·The American Journal of Forensic Medicine and Pathology·Papanu Suttipasit

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