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Sampling and laboratory investigation

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Abstract

This chapter provides a practical guide to performing, and interpreting the results of, a range of sampling techniques. The key techniques covered are synovial fluid collection and analysis, serological analysis, synovial membrane and periarticular tissue biopsy, bone biopsy and muscle biopsy.

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Figures

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4.1 Sites for arthrocentesis. Designed and drawn by Vicki Martin Design and printed with their permission (Reproduced from BSAVA Manual of Canine and Feline Clinical Pathology, 2nd edition)
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4.2 Routine techniques for the preparation of a synovial fluid smear. (a) ‘Crush-prep’ technique. (b) ‘Blood smear’ technique, which may be useful for synovial fluid with poor viscosity. Drawn by S.J. Elmhurst BA Hons (www.livingart.org.uk) and reproduced with her permission.
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4.3 If smears are not quickly air dried, this can result in significant drying artefact and loss of cell detail. These are neutrophils (arrowed) in which drying artefact has resulted in cell shrinkage; such dark, rounded up cells may be mistaken for lymphocytes. (Wright–Giemsa stain, original magnification X400)
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4.4 A blood culture medium bottle.
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4.6 Smears from synovial fluid often have a granular pink proteinaceous background. The cells are vacuolated mononuclear cells consistent with macrophages from a joint with degenerative joint disease. (Wright–Giemsa stain, original magnification X)
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4.7 Inflammatory arthropathy. Neutrophils line up in rows indicating normal viscosity of the sample, a phenomenon known as ‘windrowing’. (Wright–Giemsa stain, original magnification X)
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4.8 (a) A macrophage containing phagocytosed erythrocytes (erythrophagia). This can be pathological or can occur , in samples with iatrogenic haemorrhage. (Wright–Giemsa stain, original magnification X1000) (b) Joint with chronic haemarthrosis. Two macrophages contain intracytoplasmic haemoglobin breakdown products that include haemosiderin (dark green pigment, white arrow) and haematoidin (bright yellow pigment, black arrow). (Wright–Giemsa stain, original magnification X)
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4.9 Arrow indicates a large mononuclear cell consistent with a macrophage that contains expanded vacuolated cytoplasm in a joint with degenerative joint disease. (Wright–Giemsa stain, original magnification X)
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4.10 Neutrophils (arrowed) are the dominant cell type in an inflammatory arthropathy of either infectious or non-infectious aetiology. (Wright–Giemsa stain, original magnification X)
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4.11 Synovial fluid from a septic joint. Intracytoplasmic bacteria are identified within the degenerate neutrophils (arrowed).(Wright–Giemsa stain, original magnification X400)
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4.12 Fine-needle aspirate of a periarticular mass reveals neoplastic cells typical of a histiocytic sarcoma. (Wright–Giemsa stain, original magnification X400)
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4.13 A clump of synovial cells that have been incidentally sampled during arthrocentesis. This must not be misinterpreted as being neoplastic. (Wright–Giemsa stain, original magnification X400)
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4.14 Biopsy specimen of a histiocytic sarcoma demonstrating the extensive necrosis often present in malignant tumours. The larger the biopsy specimen, the more likely it is to be diagnostic. (Haematoxylin and eosin stain, original magnification X40)
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4.15 Biopsy specimen of synovium: there is a lymphoplasmacytic to lymphonodular infiltrate (arrowed) accompanied by synovial villous hyperplasia. (Haematoxylin and eosin stain, original magnification X40)
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4.16 A non-specific lymphoplasmacytic infiltrate is common in most inflammatory arthropathies. The arrow points to a plasma cell. (Haematoxylin and eosin stain, original magnification X400)
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4.17 Correct placement of the needle is important to maximize the chance of a diagnosis. Reactive periosteal and endosteal bone needs to be avoided. L = lesion; Rb = reactive bone. Drawn by S.J. Elmhurst BA Hons (www.livingart.org.uk) and reproduced with her permission. (Redrawn after Simon Scurrell)
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4.18 A Jamshidi needle.
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4.19 Surgical kit for performing a bone biopsy.
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4.20 Performing a bone biopsy using a Jamshidi needle.
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4.21 The core biopsy specimen is ejected in reverse through the handle.
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4.22 (a) Good quality trucut biopsy of an osteolytic lesion. (Haematoxylin and eosin stain, original magnification X20) (b) Osteosarcoma. The neoplastic cells are producing osteoid (arrowed), the hallmark of an osteosarcoma. (Haematoxylin and eosin stain, original magnification X200)
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4.23 Fine-needle aspirate of an osteosarcoma. The neoplastic cells are interspersed with bright pink matrix, consistent with osteoid. (Wright–Giemsa stain, original magnification X)
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4.24 (a) A biopsy sample taken from the thickened synovium of a stifle joint in a cat reveals granulomatous inflammation. Mycobacterial infection is a very important differential diagnosis in such a case and an acid-fast stain is required to screen for the bacteria. (Haematoxylin and eosin stain, original magnification X) (b) A Ziehl–Neelsen stain confirms the presence of acid-fast bacilli (arrowed), typical of spp. (Haematoxylin and eosin stain, original magnification X)
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4.25 (a) A biopsy sample taken from a periarticular mass in a dog indicates a malignant neoplasm, consistent with a histiocytic sarcoma. The arrows indicate mitotic figures. Immunohistochemistry can then be performed to support the diagnosis. (Haematoxylin and eosin stain, original magnification X) (b) Immunohistochemistry for the leucocyte antigen CD18 is performed and immunopositivity is indicated by the brown staining. This together with the haematoxylin and eosin findings, is typical of a histiocytic sarcoma. (Haematoxylin and eosin stain, original magnification X)

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