Laser Microdissection
Friday, June 26, 2009 9:00 am - 5:00 pm
BioPark Hertfordshire
Broadwater Road
Welwyn Garden City, Hertfordshire AL7 3AX
United Kingdom
The agenda for this event is under construction. If you have any queries or are interested in speaking at this event please contact meetingsproducer@euroscicon.com
This meeting has CPD approval
Talks include
Laser Microdissection in Parasitology - Professor Rory Post - Department of Infectious and Tropical Diseases, London School of Hygiene & Tropical Medicine
Laser Capture Microdissection of Mouse Cortex - Mary Antypa - University College London
Automatic detection of spermatozoa for laser capture microdissection - Mado Vandewoestyne,, Laboratory of Pharmaceutical Biotechnology, Ghent University , Belgium
In sexual assault crimes, laser capture microdissection (LCM) enables separation of spermatozoa and epithelial cells. We describe an automated screening method to detect spermatozoa stained with Sperm HY-LITERTM, a fluorescent kit for the detection of human spermatozoa. Different ratios of spermatozoa and epithelial cells as well as real postcoital samples were used to assess the automatic detection method. Detected spermatozoa were isolated using LCM and DNA analysis was performed. Robust DNA profiles without allelic dropout could be obtained from as little as 30 spermatozoa recovered from postcoital samples, showing that the staining and LCM had no significant influence on DNA recovery.
Additional Speakers
- Dr Roman Gonitel - Medical and Molecular Genetics, King's College London
About the Meeting chair
Dr David Whitehouse
David is an experienced academic and commercial scientist. He has more than 20 years research experience in the university sector, mostly with the MRC Human Biochemical Genetics Unit in UCL focusing on protein detection, human molecular genetics and genomics and the development of rapid diagnostic tests using monoclonal antibodies. In 2000 he transferred to the commercial sector where he specialized in the development of optical and electrophoretic devices for microbial detection and new approaches to DNA based diagnostics. He is an experienced freelance manager of intellectual property including patent applications in the biotechnological and neurosciences fields. He writes, lectures, and devises presentations and learning modules in biotechnology and healthcare for the commercial and higher education sectors.
About the Speakers
Mado Vandewoestyne obtained her Master degree in Pharmaceutical Sciences at Ghent University in 2006. She is currently working as a PhD student in the Laboratory of Pharmaceutical Biotechnology. This laboratory, where Mado is one of the quality assurance managers, performs human identification by the use of DNA profiling techniques, mainly for the Belgian Department of Justice. Mado’s main research interest is the application of laser microdissection on forensic mixed samples, resulting in pure cell preparations for DNA analysis.
Posters
AUTOMATIC DETECTION OF SPERMATOZOA FOR LASER CAPTURE MICRODISSECTION
M Vandewoestyne, Van Hoofstat D, Van Nieuwerburgh F, Deforce D
Laboratory of Pharmaceutical Biotechnology
Ghent University
Harelbekestraat 72
9000 Ghent
Belgium
Phone: +32 (0)9 264 81 39
Fax: +32(0)9 220 66 88
Mail: Mado.Vandewoestyne@Ugent.be
In sexual assault crimes, differential extraction of spermatozoa from vaginal swab smears is often ineffective, especially when only a few spermatozoa are present in an overwhelming amount of epithelial cells. Laser capture microdissection (LCM) enables the precise separation of spermatozoa and epithelial cells. However, standard sperm staining techniques are non-specific and rely on sperm morphology for identification. Moreover, manual screening of the microscope slides is time-consuming and labour-intensive.
In the present study, we describe an automated screening method to detect spermatozoa stained with Sperm HY-LITERTM (Independent Forensics, Hillside, IL), a fluorescent kit for the detection of human spermatozoa which does not rely on morphological characteristics or non-specific staining for identification. The spermatozoa are detected using an Alexa Fluor 488 derivatized mouse monoclonal antibody against proteins contained in the human sperm heads. In addition, a 4’,6-diamidino-2-phenylindole (DAPI) staining is used to detect all nuclei present on the slide. Processed slides can be viewed at low magnification, greatly increasing the speed of sperm identification.
Different ratios of spermatozoa and epithelial cells were used to assess the automatic detection method. In addition, real postcoital samples were also screened. Detected spermatozoa were isolated using LCM and DNA analysis was performed. Robust DNA profiles without allelic dropout could be obtained from as little as 30 spermatozoa recovered from postcoital samples, showing that the staining and laser capture microdissection had no significant influence on DNA recovery.
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Euroscicon
BioPark Hertfordshire
Broadwater Road
Welwyn Garden City,
Hertfordshire AL7 3AX
United Kingdom
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