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Diagnosis of Hematodinium infection in C. sapidus has previously relied on microscopic observation of fixed and stained (hematoxylin and eosin/giemsa) hemolymph samples and tissue sections for the presence of parasites (Messick 1994, Messick & Shields 2000, Infected C. sapidus were identified using smears of fresh hemolymph mixed with neutral red, as described by Stentiford and Shields (2005). Briefly, approximately 1 mL of crab hemolymph was removed from the fifth walking leg using a sterile syringe and a 27 ga. needle. Ethanol (70%) was used to sterilize the surface of the crab prior to removal of hemolymph. An equal volume of neutral red (0.04% (w/v) in 1 x phosphate buffered saline (PBS) was mixed with a subsample of hemolymph, and the sample placed on a glass slide and viewed under light microscopy to assess the presence of parasites. Source
Tags : hematoxylin and eosin, phosphate buffered saline pbs, phosphate buffered saline, sterile syringe, microscopic observation, light microscopy, neutral red, glass slide, blue crab, tissue sections, reece, ethanol, parasites, shields, pbs
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