Publication Abstract

Laser Microdissection (LMD): A new tool for marine molecular parasitology

H.J. Small, J. Sturve, G.D. Stentiford*, J.P. Bignell*, R. Hicks*, M. Longshaw*, B.P. Lyons*, S.W. Feist*

Laser-assisted microdissection (LMD) has been developed to accurately isolate distinct cell populations from a heterogeneous tissue section, cytological preparation, or live cell sample. Downstream applications typically include gene expression studies using real-time PCR and array platforms, diagnostic PCR, and protein expression studies. LMD techniques are now commonplace in mainstream biological research and clearly have suitable applications in the field of aquatic parasitology. The present study describes, as an example, the use of LMD to isolate two dinoflagellate parasites (Hematodinium spp.) from formalin-fixed wax-embedded tissue sections from two crustacean hosts. DNA was isolated from LMD parasite preparations, and partial regions (up to 300 bp) of the small subunit (SSU) and the first internal transcribed spacer (ITS1) region of the ribosomal RNA (rRNA) gene complex from the Hematodinium spp. were PCR amplified using diagnostic primers. The amplification products were sequenced to confirm DNA stability. The techniques, applications, and limitations of LMD to address questions in aquatic molecular parasitology are discussed.

Reference:

H.J. Small, J. Sturve, G.D. Stentiford*, J.P. Bignell*, R. Hicks*, M. Longshaw*, B.P. Lyons*, S.W. Feist* (2008) Laser Microdissection (LMD): A new tool for marine molecular parasitology. Diseases of Aquatic Organisms, Vol 82: 151-156

http://www.int-res.com/articles/dao2008/82/d082p151.pdf