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Diagnostic Cytology

Diagnostic Cytology

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Diagnostic Cytology
 
Diagnostic cytology involves the examination of cells which have either naturallyexfoliated or been artificially removed from a body cavity or a tissue mass. Thecytological interpretation is often valuable in establishing a diagnosis, identifyingthe disease process, directing therapy, forming a prognosis, and/or determiningwhat diagnostic procedure should next be performed.Some of the major advantages of cytological procedures are that they requireminimal equipment, have rapid turn around time and most are minimally invasiveand minimally stressful for the patient. However, since cytologic interpretation isbased upon the distinct characteristics of individual cells on a smear that lacksthe source tissue's architectural patterns, it should be mainly utilized as a rapidscreening procedure. Histopathology is usually more diagnostic and definitive inthat it allows study of both structure and form of a tissue or organ. Cytologyshould be considered a tentative diagnosis requiring a histologic confirmation,particularly in lesions with suspected neoplastic characteristics. 
10.1 METHODS OF SPECIMEN ATTAINMENTA. Solid Tissue Cytology
The particular method used for obtaining cytological specimens is determined bythe characteristics of the lesion to be sampled. The major goal is to obtain asignificant number of well-stained intact cells that reflect the composition of thelesion.
1. Touch impressions or imprints
Imprints can be prepared from excised external lesions on the living animal or from tissues removed during surgery or necropsy. The imprints are easy toobtain but they collect fewer cells than scrapings and contain greater contamination
(bacterial and cellular)
than aspirates. Therefore, imprints fromsuperficial lesions often only reflect a secondary bacterial infection and/or 
 
inflammation-induced tissue dysplasia. This markedly hinders their use indiagnosis of neoplasia. To reduce contamination and to obtain a morerepresentative sample, touch imprints are made on freshly excised surfaces. If the excised surface is very bloody or moist, it should be blotted with absorbentpaper. Touch the slide gently to the specimen, remove and quickly air dry.Dragging of the slide across the specimen will distort the cells. Tissue rich inconnective tissue imprints poorly.
2. Scrapings
Scraping has the advantage of collecting many cells from the tissue, andtherefore, is advantageous when the lesion is firm and yields few cells.Disadvantages are similar to those described for tissue imprints. The surface of the lesion is freshly excised and scraped with a scalpel blade until materialappears on the blade's surface. This material is then smeared thinly across aslide.
3. Swabs
Swab smears are collected only when imprints, scrapings and aspirates cannotbe made; e.g., fistulous tracts, nasal and vaginal collections. The lesion or tissueis swabbed with a moist, sterile cotton swab. Sterile isotonic fluid, such as
0.9%NaCl
, should be used to moisten the swab. This helps minimize cell damageduring sample collection and smear preparation. After sample collection, theswab is gently rolled, not rubbed, along the flat surface of a clean glassmicroscope slide.
4. Fine Needle Aspirations
Fine needle aspiration biopsies can be collected from masses such as lymphnodes, nodular lesions and internal organs. Aspiration of cutaneous massesavoids the superficial contamination common with imprints or scrapings, butcollects fewer cells than scrapings. The tissue to be aspirated is immobilized asclose to the skin surface as possible.
A 21-25
gauge needle attached to a 3-20ml syringe is used
(a 12-ml syringe is a good all-around size)
. The softer thetissue being aspirated, the smaller the needle and syringe used. When needleslarger than 21 gauge are used, tissue cores tend to be aspirated, resulting in apoor yield of free cells suitable for cytological preparation. Also, larger needlestend to increase the incidence of blood contamination.The needle with syringe attached is introduced into the center of the mass andstrong negative pressure is applied The cell population retrieved from a mass ishighly dependent upon the site aspirated. In order to obtain a representativesample, the needle is redirected and moved to several areas in the mass, takingprecautions to prevent the needle from leaving the mass. Negative pressure ismaintained during the redirection. However, when the mass is not large enough
 
for the needle to be redirected and moved without danger of the needle's leavingthe mass, negative pressure is relieved during redirection and movement of theneedle. In this situation, negative pressure is applied only when the needle isstatic. In both cases, aspirate may or may not appear in the syringe, but theneedle will contain sufficient tissue for smears. Before removing the needle fromthe mass, the negative pressure is relieved. The needle is removed from themass and skin and the needle contents are expressed onto a slide and a smear prepared.
10.1 Fluid Cytology1. Centesis
Centesis is the process of perforation or tapping as with an aspirator, trocar, or needle, to obtain abdominal, thoracic, synovial or cerebrospinal fluid. The size of the trocar or needle used is dependent upon the species and centesis site.Whenever possible, at least 3 ml of fluid should be aseptically obtained toprovide an amount sufficient for determination of specific and concentration for microscopic examination. Since exudates coagulate and modified transudatesmay coagulate, a portion of the specimen should be collected in an
EDTA
tube.A sterile tube should be used for one portion to assure a satisfactory specimen inthe event that culture is indicated. Direct and sediment smears should be madefor cytological examination. Sediment smears are prepared by centrifu-gation of the fluid for 5 minutes at 165-360 G to concentrate the cells. In addition tocytological examination, other properties
(e.g., specific gravity, total protein,glucose)
should be determined in order to pin-point the pathogenesis of theoccurrence.
2. Catheterization
Catheterization procedures are used for transtracheal/ bronchial and prostaticwashes. Once obtained, the fluid specimens are handled in a manner similar tocentesis fluids.
10.2 PREPARATION OF SMEARS10.2.1 Preparation of smears from aspirates of solid masses
Several methods can be used to prepare smears for cytologic evaluation of solidmasses, including lymph nodes. The experience of the person preparing thesmears and characteristics of the sample influence the choice of smear preparation technique. No matter what the technique used, the resultant smear must have a thin area where cells may settle flatly and expose a large surfacearea to view. If the smear is thick, the cells are supported more upright on theslide and have a smaller diameter. Cellsin thick areas are taller and thus staindarker. In addition, thick layers of proteinaceous and necrotic debris in the dried

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