Laser-assisted microdissection : a new tool for aquatic molecular parasitology

Laser-assisted microdissection (LMD) has been developed to isolate distinct cell populations from heterogeneous tissue sections, cytological preparations, or live cell samples. Downstream applications typically include gene expression studies using real-time PCR and array platforms, diagnostic PCR,...

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Bibliographic Details
Published in:Diseases of aquatic organisms 2008-11, Vol.82 (2), p.151-156
Main Authors: SMALL, H. J, STURVE, J, BIGNELL, J. P, LONGSHAW, M, LYONS, B. P, HICKS, R, FEIST, S. W, STENTIFORD, G. D
Format: Article
Language:English
Subjects:
Animal aquaculture
Animal productions
Animals
Biological and medical sciences
Biological Sciences
Biologiska vetenskaper
Brachyura
Brachyura - parasitology
Cancer pagurus
cytology
Dinoflagellida
Dinoflagellida - cytology
DNA
DNA, Intergenic - isolation & purification
DNA, Protozoan - isolation & purification
Fundamental and applied biological sciences. Psychology
General aspects
Hematodinium
Hepatopancreas
Hepatopancreas - parasitology
instrumentation
Intergenic
isolation & purification
Lasers
Marine
methods
Microdissection
Microdissection - instrumentation
Microdissection - methods
Muscle
Muscle, Skeletal - parasitology
parasitology
Polymerase Chain Reaction
Portunus trituberculatus
Protozoan
Skeletal
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Summary:Laser-assisted microdissection (LMD) has been developed to isolate distinct cell populations from heterogeneous tissue sections, cytological preparations, or live cell samples. 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 pathology and parasitology. The present study used LMD to isolate 2 dinoflagellate parasites (Hematodinium spp.) from formalin-fixed paraffin-embedded tissue sections from 2 crustacean hosts, Cancer pagurus and Portunus trituberculatus. DNA was isolated from LMD parasite preparations, and partial regions (up to 300 bp) of the small subunit and the first internal transcribed spacer region of the rRNA gene complex from the Hematodinium spp. were PCR amplified using diagnostic primers. The amplification products were sequenced to confirm the identity of the targeted regions. The techniques, applications, and limitations of LMD to address questions in aquatic molecular pathology and parasitology are discussed.
ISSN:0177-5103
1616-1580
DOI:10.3354/dao01983