Cytology

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XQWCat
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251872
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Cytology
Updated:
2013-12-08 22:52:03
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Clin Path
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Clin,Path
Description:
Cytology in veterinary clinical pathology
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  1. Mast cell tumor
    • diagnosed with cytology, no need for biopsy.
    • Presents as "disease" of allergy, doesn't go down in 24h with treatment, look for mast cell tumor (systemic histamine)
    • Usually pink, raised epidermal mass, can look like anything.  Give benadryl pre-op before removing.
  2. Cytology
    study of cells. Formation, structure, function
  3. histopathology
    study of tissues.  Cells, but in relationship to tissues
  4. hypertrophy
    increase in cell SIZE and/or FUNCTIONAL ACTIVITY in response to a stimulus (hypertrophic cardiomyopathy)
  5. hyperplasia
    increase in cell NUMBERS in response to a pathologic or physiologic stimulus (skin callus)
  6. neoplasia
    increase in cell growth and multiplication INDEPENDANT of stimulus (sarcoma, leukemia, carcinoma, etc)
  7. dysplasia
    abnormal development (hip dysplasia)
  8. metaplasia
    • substitution of one cell type for another adapted from a stimulus (columnar cells in lining of bronchi--replaced by scar due to chronic inflammation)
    • irritation inflammation altersation of cell signal (hormonal imbalance) leading to tissue restructure
  9. Uses of cytology
    • diagnoses
    • differentiating disease processes (inflammation vs. neoplasia)
    • directing therapy (otitis externa, yeast or bacteria, check progress)
  10. Cytologic specimens can be obtained by
    • imprints (press on)
    • swabs (ear, vaginal)
    • skin scrape (mites)
    • fine needle aspirate
    • smear preps for solid masses (squash prep, modified compression smear, starfish prep)
  11. preparation of cytologic samples, check before sending out for
    • # of cells
    • intact vs damaged
    • stain appropriate
    • 3-4 samples if possible
  12. Microscope eval of cytology
    • ID cell type and disease process
    • cell morphology, low power: all areas stained, localized increased cellularity
    • cell morphology, high power: characterize cellularity and composition (types and relative numbers, compare individual cells)
    • Oil immersion: ID specific nuclear criteria of malignancy and inflammatory
  13. 5 categories of interpretation of cytology
    • inflammation: presence of neutrophils, often blood contamination (hard to tell)
    • cyst formation: large numbers of mature, keratinized, squamous epithelial cells or amorphous whisps with low cellularity
    • hemorrhagic lesion: blood and macrophages, some eating RBC or hemosiderin (differentiate from blood contamination)
    • neoplasia: homogenous population
    • mixed cell population: both inflammatory (neutrophils, macrophages, etc) and noninflammatory (epithelial ro mesenchymal)
  14. hemosiderin
    iron storage complex in cells (brown or black)
  15. interpretation of inflammatory cytology, 5 categories
    • suppurative (purulent)
    • granulomatous
    • pyogranulomatous
    • eosinophils
    • lymphoplasmacytic
  16. interpretation of inflammatory cytology
    • presence of WBCs (normal response to tissue damage or invaders) due to chemotactic effect. Neutro first, then macrophages, sometimes eosinophils/lymphocytes
    • Can be different types of inflammation (% neutrophils, other cell types, may help determine cause).  5 types.
  17. Suppurative inflammation cytology
    purulent.  Greater than 85% neutrophils
  18. Granulomatous inflammation cytology
    greater that 15% macrophages and FEW neutrophils
  19. pyogranulomatous inflammation cytology
    greater than 15% macrophages and MANY neutrophils
  20. eosinophilic inflammation cytology
    greater than 10% eosinophils with INCREASED neutrophils
  21. lymphoplasmacytic inflammation cytology
    • large proportion of mature lymphocytes (small lymphs and plasma cells)
    • often seen in cats, esp in GI tract.
  22. septic inflammation cytology
    • degenerative neutrophils, water in cell, nuclear swelling.  
    • Infection, usually bacterial.  Can see intracellular/phagocytized bacteria.  
    • Derivative of supporative inflammation
    • Most often seen in fluid cytology, ascites or pleural effusion
  23. cyst cytology interpretation
    • Lots of mature, keratinized squamous epithelial cells or amorphous material with few cells. (boring cytology)
    • 3 types of cutaneous cysts depending on what gland has duct expansion: epidermal inclusion (hair follicle), apocrine (sweat), sebaceous (thick, cheesy material on skin, often allergic)
  24. Hemorrhagic cytology interpretation
    Very hard to avoid contamination, not hemorrhagic unless has hemosiderin pigment or whole RBCs within macrophages
  25. benign neoplastic cytology interpretation
    • homogenous population of single cell type
    • Hyperplasia without signs of malignancy
    • Uniform population of cells, uniform in cytoplasmic and nuclear size, shape, and nuclear/cytoplasmic ratio (N:C)
    • nucleoli may be present, consistent between cells
  26. malignant neoplastic cytology interpretation
    • homogenous population of a single cell type
    • must have at least 3 abnormal nuclear configurations
    • ID primary cell type involved
  27. Criteria of malignancy in neoplastic cytology
    • nuclear features: anisokaryosis, pleomorphism, macrokaryosis, high/variable N:C ratio, increased mitotic activity, coarse chromatin pattern, nuclear modeling, multinucleation, nucleoli that vary in size, shape and number
    • cytoplasmic features: increased basophilia, abnormal vacuoles/granuoles, cytophagia
  28. anisokaryosis
    • variation in nuclear size between cells (1.5x or greater)
    • criteria of malignancy
  29. pleomorphism
    • variation in nuclear and cell size and shape
    • criteria of malignancy
  30. high nuclear to cytoplasmic ratio (N:C)
    • nucleus comprises more than 1/2 total cell size
    • criteria of malignancy
  31. mitotic figures
    • organization and separation fo nuclear chromatin (ropy chromatin pattern)
    • criteria of malignancy
  32. large or irregularly shaped nucleoli
    • prominent nucleoli of variable size, shape and number between cells.  Angular nucleoli or nucleoli larger than mature erythrocytes suggest malignancy
    • criteria of malignancy
  33. coarse or clumped chromatin
    • nuclei with clearly visible light (euchromatin) and dark (heterochromatin) areas indicate increased cell activity
    • criteria of malignancy
  34. nuclear molding
    • nuclei which distort or compress adjacent nuclei indicate a cell population with uncontrolled proliferation and lack of contact inhibition
    • criteria of malignancy
  35. multinucleation
    • multiple nuclei seen in malignant cell population
    • criteria of malignancy
  36. epithelial tumor cytology
    exfoliate in clusters and sheets, round to polygonal cells, well-defined cell borders (adenomas, carcinomas)
  37. Four categories of tumor type
    • epithelial
    • mesenchymal
    • round cell
    • neuroendocrine
  38. mesenchymal tumor cytology
    • do not exfoliate well (hard to get cytology)
    • individual elongated or spindle-shaped cells (fibrous tissue)
    • sarcomas
  39. Round cell tumor cytology
    • exfoliate well and appear as separate cells (less sheets)
    • round cells with well defined cytoplasmic margins
    • MCT, lymphoma, plasmocytoma, histiocytoma, transmissable venereal tumor (often on nose)
  40. neuroendocrine tumor cytology
    islands or sheets of cells with round to oval stippled nucleus with scant pink cytoplasm
  41. mixed cell population cytology interpretation
    • preparation has both inflammatory and noninflammatory cells.
    • Tumors associated with inflammation may include squamous cell carcinomas, nasal tumors and bladder tumors.  
    • Locally aggressive, lots of inflammation induced by tumor
  42. Fluid cytology
    • smears prepared immediately after fluid collection (denatures over time)
    • Collect in EDTA tubes (blood contamination)
    • centrifuge on urine setting, remove supernatant, make smear from sediment
  43. cerebrospinal fluids cytology
    • evaluate neurologic disorders
    • collect from cisterna magna or lumbosacral space
    • need spinal needle with stylet, clippers, EDTA tubes, STERILE gloves and drape
  44. Preparation/procedure for Cerebrospinal fluid cytology
    • general anesthesia so no movement
    • intubation
    • sterile prep of procedure site
    • Postion, right lateral recumbency for cisternal magna (high tap) and ventral recumbency with flexed coxofemoral joints for low tap
  45. results for cerebrospinal fluid cytology
    • Normal findings are no RBC, should be clear, < 25 nucleated cells/mL, most lymphocytes, low protein,
    • CPK - Low without damage. Ancillary test for nonspecific damage to neural tissue, increased levels after seizures.
  46. Centesis
    • Introduction of a needle into a body cavity or organ to remove fluid
    • abdominocentesis, thoracocentesis, cystocentesis, arthrocentesis
  47. equipment for abdominocentesis/thoracocentesis
    • needle of sufficient length
    • extension set
    • 3-way stopcock
    • 20-60ml syringe
    • clippers
    • scrub solution
    • bowl for fluid (when syringe is full)
    • sample tubes and aerobic/anaerobic culturettes
  48. Collection of abdominocentesis/thoracocentesis
    • sedation not required
    • aseptic preparation of site
    • Thoraco: sternal position (fluid sinks to bottom, air rises to top), 7-8 intercostal space along cranial aspect (avoid nerve/vessel)
    • abdomino: standing, lateral or dorsal recumbancy (ultrasound), needle into ventral abdomen to right of midline, 1-2 cm caudal to umbilicus
  49. results of abdominocentesis or thoracocentesis
    • abdomino: colorless and transparent, no odor, low cell counts
    • thoraco: clear and colorless to slightly yellow, low cellularity, low protein
    • small fluid not necessarily pathologic
    • Look at color, turbidity, cell counts, types and morphology, EDTA tubes for cell count and cytological exam, refractometry for protein, red tops for clinical chem and protein
  50. Types of thoraco or abdomino effusions (6)
    • exudates
    • transudates
    • modified transudates
    • chylous effusion
    • hemorrhagic fluid
    • FIP
  51. exudates in abdominal or thoracic fluid effusions
    • fluids with increased cellularity  and increased protein concentration.  
    • Septic has degenerative neutro and bacteria in macrophage
    • nonseptic
    • both are inflammation, neutrophils and cloudy solution
  52. transudates in abdominal or thoracic fluid effusions
    • Fluids with low cellularity and low protein concentration.  
    • Hypoproteinemia (low albumin causes 3rd spacing)
    • early right CHF
  53. Modified transudate in abdominal or thoracic fluid effusions
    • Low to moderate total cell counts and high total protein
    • almost anything can cause.  neoplasia, cardiac disease, FIP
  54. Chylous effusion in abdominal or thoracic fluid effusions
    • milky fluid due to leakage of chyle (increased chylomicrons).
    • Rupture of thoracic duct, cardiomyopathy, neoplasia, heartworm disease, trauma, rupture of lymphatic vessel, lung lobe torsion, etc.
  55. Hemorrhagic fluid in abdominal or thoracic fluid effusions
    • blood contamination?
    • acute hemorrhage: PLATELETS, no erythrophagocytosis (too soon)
    • few hours old: macrophages with erythrophagia
    • few days old: macrophages with hemosiderin
    • trauma, neoplasia
    • pericardial effusions: neoplasia, idiopathic
  56. FIP in abdominal or thoracic fluid effusions
    • "straw colored" fluid
    • modified transudate or exudate based on cell count (never transudate)
    • protein > 3.5 g/dL 
    • non-degenerative neutrophils, macrophages, lymphocytes, protein precipitate
    • indirect fluorescence assay to test for coronavirus
  57. arthrocentesis
    • polyarithritis, immune or tick-borne
    • adequate sedation, clip and scrub, ASEPTIC, need sterile gloves, 1-1 1/2 22-25G needles, 3mL syringe, glass slides, EDTA tube, culturette, diff-quick stain
    • Go through capsule, try not to hit cartilage
  58. arthrocentesis procedure
    • palpate bony landmarks to find insertion point between bones
    • attach needle to syringe and advance needle into joint space ("pop")
    • aspirate with gentle suction
    • 3-4 samples from normal and abnormal joints
  59. arthrocentesis results (normal)
    • normal joint fluid is sticky, few erythrocytes, 90% mononuclear cells and 5% neutrophils with "rowing"
    • Look at volume, color (clear to slight yellow), turbidity (should be clear), viscosity (hyaluronic acid), cellularity, cell distribution (rowing due to viscosity), protein concentration with refractometer
  60. arthrocentesis results, abnormal, reduced viscosity
    • bacterial inflammation
    • effusion as a result of diluted hyaluronic acid.
    • Puffy joint = bacteria
  61. arthrocentesis results, abnormal, inflammatory
    • infectious and noninfectious (lyme, immune)
    • cell count > 10,000 cells/uL
    • mostly neutrophils
    • mononuclear increased
    • protein increased
    • culture recommended
  62. arthrocentesis results, abnormal, degenerative
    • cells < 5000 cells/uL
    • usu mononuclear predominate
    • neutrophils and protein normal to increased
  63. Tracheal wash
    • Chronic cough or bronchoalveolar disease
    • Orotracheal (through mouth, easily contaminated), nasotracheal (through nose), transtracheal (through skin, purest, put tube between tracheal rings.  Rarely done)
    • Bronchalveolar lavage (through fiberoptic bronchoscope)
  64. Transtracheal wash preparation and equipment
    • site depends on size of animal
    • clip and scrub site
    • sedatation often not required
    • local with lido
    • sit or sternal recumbancy with head up
    • "through the needle" catheter with needle guard, lidocaine, hemostat, sterile gloces, syringes with non-bacteriostatic saline
  65. transtracheal wash procedure
    • needle inserted bevel down into trachea until "pop" (cough or swallow)
    • catheter advanced through needle, withdraw needle, use needle guard.
    • stabilize with hemostat, inject warm sterile saline
    • aspirate fluid back into syringe
    • repeat until mucoid or purulent material are recovered
    • red top and EDTA tube, make smears and cultures
    • LIGHT sedation so they can cough/swallow
  66. transtracheal wash results
    • normal: few cells, small mucus, epithelial cells primarily (columnar ciliated or nonciliated cuboidal), neutrophils, lymphocytes, eosinophils, plasma cells, mast cells and erythrocytes rarely, alveolar macrophages most common
    • abnormal: exudate, cellular with mucus, neutrophils in acute inflammation, mononuclear macrophages in chronic inflammation, maybe bacteria or fungal, eosinophils in allergic or parasitic, neoplasitc or erythrocytes possible
  67. histopathology
    • evaluates tissue architecture, observe in relation to neighboring cells
    • preparation of a sample involves several complex steps and some specialized equipment
    • first step is tissue biopsy
  68. tissue biopsy
    • several different techniques
    • samples fixed in 10% neutral phosphate-buffered formalin
    • Place tissue in formalin, use cassette for small samples
  69. punch biopsy
    • disposable punches of various sizes, may or may not need sutures.
    • Fast and easy
    • rotate punch until tissue is sectioned, pull out or flush out onto tongue depressor, let dry then put depressor into formalin jar.
  70. wedge biopsy
    • obtained with scalpel, near edge of lesion, want lesion, transitional and normal tissue in sample.
    • large and variably sized. Good for solitary lesions (mass)
  71. Toxicology
    • No "tox screen", need history to know what to look for.
    • contact lab for appropriate procedure.  
    • Can be whole blood, serum, vomit, gastric lavage, feces, urine in live animal
    • whole blood, serum, urine, gut contents, organs or tissues in necropsy
  72. Things to ask yourself in cytology
    • is sample diagnostic?
    • Homogenous or heterogenous?
    • inflammation?
    • round, epithelial, azynchymal, neuroendocrine? 
    • criteria of malignancy

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