This power point provides basic information on various cytological aspects in tumour diagnosis

1. Assignment on
Fine Needle
Aspiration Cytology
KARTHIKA SKARTHIKA S
M-5609M-5609
VPL – 606VPL – 606
Department of Veterinary Pathology
Indian Veterinary Research Institute

2. INTRODUCTION
Cytopathology refers to diagnostic techniques that are used toCytopathology refers to diagnostic techniques that are used to
examine from various body sites toexamine from various body sites to
determine the and of disease.determine the and of disease.
cells
causenature
(Sr. Kamenica 2002)
Cytology refers to the microscopic evaluation of cells, veryCytology refers to the microscopic evaluation of cells, very
useful in the clinical diagnosis of neoplasia.useful in the clinical diagnosis of neoplasia.
( Wellman ,1990)

3. Cont.. George Papanicolaou claimed to be the father of clinicalGeorge Papanicolaou claimed to be the father of clinical
cytology.cytology.
 Cytology is a joint effort between the clinician andCytology is a joint effort between the clinician and
cytologist.cytologist.
 General principles of cytology is identify inflammation vs.General principles of cytology is identify inflammation vs.
neoplasia of Skin, Lymph nodes, Bone marrow and otherneoplasia of Skin, Lymph nodes, Bone marrow and other
vital organs.vital organs.
 Also examine Fluids, e.g. thoracic, abdomen, synovial fluidAlso examine Fluids, e.g. thoracic, abdomen, synovial fluid
 Staging of estrous cycle in bitches (vaginal cytology)Staging of estrous cycle in bitches (vaginal cytology)

4. Indications For CytologicalIndications For Cytological
ExaminationExamination
 Cutaneous or subcutaneous masses- peripheral lymphadenopathy (fine needleCutaneous or subcutaneous masses- peripheral lymphadenopathy (fine needle
aspiration or imprint cytology)aspiration or imprint cytology)
 Intra-abdominal or intrathoracic masses (fine needle aspiration under ultrasoundIntra-abdominal or intrathoracic masses (fine needle aspiration under ultrasound
guidance)guidance)
 Haematological diseases (bone marrow aspiration)Haematological diseases (bone marrow aspiration)
 Examination of body cavity fluids (pleural, peritoneal and pericardial effusions)Examination of body cavity fluids (pleural, peritoneal and pericardial effusions)
 Examination of urine sediment (e.g. stained preparation in addition to routine wetExamination of urine sediment (e.g. stained preparation in addition to routine wet
prep to check for neoplastic cells)prep to check for neoplastic cells)
 Joint disease (joint tap to examine synovial fluid)Joint disease (joint tap to examine synovial fluid)
 Neurological disease (cerebrospinal fluid analysis)Neurological disease (cerebrospinal fluid analysis)
 Respiratory disease (tracheal wash, bronchoalveolar lavage, and lung aspirate)Respiratory disease (tracheal wash, bronchoalveolar lavage, and lung aspirate)
 Prostatomegaly (prostatic wash or fine needle aspirate of the prostate under ultrasoundProstatomegaly (prostatic wash or fine needle aspirate of the prostate under ultrasound
guidance)guidance)
 Oestrus detection (vaginal cytology to assess stage of the oestrus cycle)Oestrus detection (vaginal cytology to assess stage of the oestrus cycle)
 Nasal disease (nasal flushings)Nasal disease (nasal flushings)
 Conjunctival and/or ocular disease (conjunctival swab, third eyelid or cornealConjunctival and/or ocular disease (conjunctival swab, third eyelid or corneal
swab/scrape)swab/scrape)

5. BIOPSYBIOPSY
Removal of a sample of tissue or cells taken from any part of the
body.
IndicationsIndications
Determine tissue diagnosis where clinical diagnosis is in doubtDetermine tissue diagnosis where clinical diagnosis is in doubt
Determine whether benign or malignantDetermine whether benign or malignant
Ascertain the degree of differentiation (grade)Ascertain the degree of differentiation (grade)
Ascertain extent of spread of disease (stage)Ascertain extent of spread of disease (stage)
Determine the tissue of originDetermine the tissue of origin
Determine different therapeutic pathwaysDetermine different therapeutic pathways
Surgical resection margins provide critical informationSurgical resection margins provide critical information

6. Types of biopsyTypes of biopsy
Cytological biopsyCytological biopsy
 FNACFNAC
 Exfoliative cytologyExfoliative cytology
 Brush cytologyBrush cytology
 Fluid spin cytologyFluid spin cytology
Histological biopsyHistological biopsy
Incisional biopsyIncisional biopsy
Core biopsyCore biopsy
CurettageCurettage
Open incisional biopsyOpen incisional biopsy
Frozen section biopsyFrozen section biopsy
Excisional biopsy/Excisional biopsy/open biopsy

7. Needle biopsy - Cells removed using a thin needle
Aspiration biopsy-Cells drawn out with a hollow needle that uses suction
Core needle biopsy -Sample of tissue is removed using a hollow core needle
that has a special cutting edge
Vacuum-assisted biopsy -A number of samples of tissue are taken using a
special rotating probe
Endoscopic biopsy -Area is viewed with an endoscope, tool is passed
through the tube to take the biopsy sample
Incisional biopsy - a portion of a mass is removed by cutting it out
Excisional biopsy - a mass is completely removed, such as a breast lump
Punch biopsy - a core of skin is removed with a special biopsy tool
Skin biopsy - a small piece of skin is cut off with a scalpel
Shave biopsy - top layers of skin are shaved off with a special blade
Bone marrow biopsy - a long needle is inserted into the bone marrow to
collect cells
Cancer liquid biopsy - Collection and evaluation of mitotic DNA
released into blood by cancerous cells

9. Punch biopsy

10. Endoscopic biopsyEndoscopic biopsy

11. Excitional and Incitional BiopsyExcitional and Incitional Biopsy

13. Renal biopsy in a cat

14. Cytopathology Methods
1.1. Exfoliative cytology – Vaginal cytology,Bronchio-alveolarExfoliative cytology – Vaginal cytology,Bronchio-alveolar
lavageslavages
2. Abrasive cytology - Scrapings2. Abrasive cytology - Scrapings
3. Fine needle aspiration cytology –3. Fine needle aspiration cytology – FNA, FNABFNA, FNAB
4. Fluid Cytology4. Fluid Cytology - Effusions from the body cavitiesEffusions from the body cavities

15. Fine needle aspiration cytology
 Examination of cells aspirated or imprinted from body cavities,Examination of cells aspirated or imprinted from body cavities,
mammary gland secretions, nasal discharges, bronchialmammary gland secretions, nasal discharges, bronchial
washings, bone marrow, lymph nodes and Deep sited organs –washings, bone marrow, lymph nodes and Deep sited organs –
guidance of CT scan, US .guidance of CT scan, US .
 The modern method of fine needle aspiration cytology wasThe modern method of fine needle aspiration cytology was
originally described by Martin and Ellis in 1930originally described by Martin and Ellis in 1930
 It yields cells rather than a tissue sample, so that it is possible toIt yields cells rather than a tissue sample, so that it is possible to
assessassess only cellular featuresonly cellular features and not the architecturaland not the architectural
characteristics of the tumour tissue. Fine-needle aspiration hascharacteristics of the tumour tissue. Fine-needle aspiration has
many positive qualities. It is relatively painless and free ofmany positive qualities. It is relatively painless and free of
complications.complications.
(Vail et al,1996)(Vail et al,1996)

16. Fine Needle Aspiration (FNA)Fine Needle Aspiration (FNA)
 The best overall method for sampling any mass or proliferativeThe best overall method for sampling any mass or proliferative
lesion, s/c glandular organs like lymph node, mammary glandlesion, s/c glandular organs like lymph node, mammary gland
or salivary glandor salivary gland
 Fine needle procedure (21 to 25 gauge)Fine needle procedure (21 to 25 gauge)
 Performed on the following tissue types:Performed on the following tissue types:
 Neck lymph nodes, cystsNeck lymph nodes, cysts
 Thyroid gland, salivary glandThyroid gland, salivary gland
 Breast tissueBreast tissue
 Any lump that can be feltAny lump that can be felt
 70-80% success rate70-80% success rate
 The four fundamental requirements on which the successThe four fundamental requirements on which the success
of FNAB depends areof FNAB depends are representativenessrepresentativeness,, adequacy of theadequacy of the
samplesample and highand high quality of preparationquality of preparation with relevant andwith relevant and
correct clinical / radiological informationcorrect clinical / radiological information

17. Methods Of Sample CollectionMethods Of Sample Collection
Aspiration TechniqueAspiration Technique
Continous suction methodContinous suction method
Intermittent suction methodIntermittent suction method
Non - Aspiration TechniqueNon - Aspiration Technique/ Capillary/ Capillary
Technique/Stab Technique – Highly vascularTechnique/Stab Technique – Highly vascular
tissue- spleen, lymphnodetissue- spleen, lymphnode

18. Selection Of Syringe And NeedleSelection Of Syringe And Needle
 22 – 25 gauge needle22 – 25 gauge needle
 3 – 20ml syringe3 – 20ml syringe
 The softer the tissue – the smaller the needle and syringe used(lymphThe softer the tissue – the smaller the needle and syringe used(lymph
nodes – 3ml)nodes – 3ml)
 The basic technique uses a 25-gauge needle and a 10-mL syringe
 Larger needles – Tissue cores aspiratedLarger needles – Tissue cores aspirated
Poor yield of free cellsPoor yield of free cells
Greater blood contaminationGreater blood contamination
 A larger bore needle and increased suction may be necessary for
mesenchymal tumours
 A 12 ml syringe is a good choice if texture is unknown (Rick L.CowellA 12 ml syringe is a good choice if texture is unknown (Rick L.Cowell
et.al, Diagnostic cytology and hematology of the dog and cat)et.al, Diagnostic cytology and hematology of the dog and cat)

19. Current Biopsy Fine NeedleCurrent Biopsy Fine Needle
Cameco Syringe Gun

20. FNA ProcedureFNA Procedure
 Preparation of the SitePreparation of the Site
 The mass is stabilised with one handThe mass is stabilised with one hand
Needle with syringe attached introduced intoNeedle with syringe attached introduced into
mass Withdraw plunger to 3/4mass Withdraw plunger to 3/4thth
volumevolume
of syringe Needle is moved back andof syringe Needle is moved back and
forth repeatedly passing through 2/3forth repeatedly passing through 2/3rdrd
diameterdiameter
of massof mass
Avoid use of US gelAvoid use of US gel
Do not allow needle to exit the mass while
negative pressure is being applied

21.  Negative pressure released Needle isNegative pressure released Needle is
removed from mass and skin Needle isremoved from mass and skin Needle is
removed from syringe Air drawn intoremoved from syringe Air drawn into
syringe Needle replaced onto the syringesyringe Needle replaced onto the syringe
and aspirate expelled onto the middle of aand aspirate expelled onto the middle of a
microscope slidemicroscope slide

22. Fine needle aspiration cytology method ( Thrall et al,1984)

24. Non – Aspiration ProcedureNon – Aspiration Procedure
 No negative pressure is appliedNo negative pressure is applied
 Barrel of syringe filled with air prior toBarrel of syringe filled with air prior to
introductionintroduction
 Needle moved back and forth in a stabbingNeedle moved back and forth in a stabbing
motionmotion
 Prevents desiccation and coagulation ofPrevents desiccation and coagulation of
samplesample
 Yield samples of equal or better qualityYield samples of equal or better quality

25. Collection TipsCollection Tips
 Make and submit multiple slidesMake and submit multiple slides
 Stain and briefly examine for cellularity –Stain and briefly examine for cellularity –
averageaverage 100 cells100 cells
 Use new needle and syringe for each massUse new needle and syringe for each mass
 Needle passages should be quick adequateNeedle passages should be quick adequate
pressurepressure
 Avoid blood dilution – prolonged aspirationAvoid blood dilution – prolonged aspiration
large needlelarge needle
A minimum of 4 -5 slides from several sites from any lesion
Any time material is visible in the hub of needle, STOP collection

26. Preparation Of SlidesPreparation Of Slides
 Slide – over – slide smears ( Squash Preps )Slide – over – slide smears ( Squash Preps )
 Blood Smear TechniqueBlood Smear Technique
Best method of preparing slide from FNB

27.  Needle spread/Starfish preparationNeedle spread/Starfish preparation

28. Preparation TipPreparation Tip
 Do not let the sample dry or clotDo not let the sample dry or clot
 Do not spray the sample from long distanceDo not spray the sample from long distance
 Avoid making too thick a smearAvoid making too thick a smear
 Do not allow cytologic specimens to come in direct contact
with ice because cell lysis will occur (do not refrigerate
smears).
 Changes occur in the fluid samples with storage, such as
phagocytosis of erythrocytes (within a few hours) and bacteria
(within 30 min), so freshly made smears should ideally be
provided along with any fluid samples

29.  Formalin-fixed cytology preparations must be
stained with either H&E or Papanicolaou stain.
 Unstained cytology slides should never beUnstained cytology slides should never be
mailed with or stored near samples in formalinmailed with or stored near samples in formalin

30. Stains
Romanowsky type stainsRomanowsky type stains
(for air dried slides)(for air dried slides)
Wright’s stainWright’s stain
Giemsa stainGiemsa stain
Wright’s Giemsa stainWright’s Giemsa stain
Diff- Quik stainDiff- Quik stain
 Cytoplasmatic features are wellCytoplasmatic features are well
preservedpreserved
 Nuclear and nucleolar featuresNuclear and nucleolar features
are less preserved.are less preserved.
Diff- Quik stainDiff- Quik stain

31. Papanicolaou
stain
 Nuclear and nucleolar featuresNuclear and nucleolar features
are better preservedare better preserved
 Cytoplasmic changes andCytoplasmic changes and
micro-organismsmicro-organisms areare
not demonstrated.not demonstrated.
 CumbersomeCumbersome
 Hematoxylin stain + Orange stainHematoxylin stain + Orange stain
+ Polychromic stain =+ Polychromic stain =
Papanicolaou stain.Papanicolaou stain.

32.  Only one colourOnly one colour
 Not a permanent stainNot a permanent stain
 Nucleoli well demonstratedNucleoli well demonstrated
 Good for a bone marrow aspiratesGood for a bone marrow aspirates
New Methylene Blue

33. Observation of different criteria's :-Observation of different criteria's :-
 Artifact during preservation & staining.Artifact during preservation & staining.
 Nuclear/ cytoplasmic ratio:-Nuclear/ cytoplasmic ratio:- Normal / increased / decreased
 Chromatin pattern and colorChromatin pattern and color Smooth / finely stippled /
hazy /coarse/ clumped / smudged
 Nucleolar appearanceNucleolar appearance
*Nuclear size : Variable / uniform
*Nuclear or cytoplasmic : Yes / No
membrane distortion
 Cytoplasmic featuresCytoplasmic features
 Extracellular matrix visiability and colorExtracellular matrix visiability and color

34. Artifacts and contaminants
SquamesGlove powder
Stain precipitate
Nuclear Debris “Ghost”cells

35. NeutrophilsNeutrophils
 Large numbers of neutrophils indicate acute
inflammation and often are accompanied by
smaller numbers of macrophages- infection or
foreign body reaction
 Truly pathogenic bacteria should be within
the cytoplasm of the phagocytic cells
Neutrophils

36.  variable morphology
 If a macrophage exhibits vacuolation or phagocytosis, it is
often described as activated.
 Can be multinucleated, especially with foreign body reactions
and in longstanding lesions.
 Under certain circumstances, they may become epithelioid-
oval or round nuclei and small, often indistinct, nucleoli. Their
cytoplasm is expanded but uniformly stained and not
vacuolated
 Common in bronchoalveolar lavage fluid, body cavity, joint
fluids
Macrophages

37.  segmented nuclei
 eosinophilic cytoplasmic granules
 slightly larger than neutrophils
 difficult to distinguish poorly stained eosinophils from neutrophils
 Allergies, superficial cutaneous viral infection in cats, eosinophilic
granuloma in horses, canine eosinophilic folliculitis and furunculosis
 Some cases of canine cutaneous mast cell tumors have a very high
proportion of eosinophils and very few mast cells
 Rabbits, guinea pigs, birds and reptiles - heterophils with eosinophilic
granules
Eosinophils

38.  small with very little cytoplasm and smudged chromatin with
no nucleoli
 The almost-round nuclei are similar in size to those of red
cells.
 Medium and large lymphocytes, which have a slightly more
open chromatin pattern and more cytoplasm, can also be seen
in inflammatory processes
 If mostly medium to large cells are present, lymphoma is a
possibility
 Normal and reactive lymphocytes in body fluids often appear
larger than the same cells from soft tissue.
Lymphocytes

39.  Multinucleated Cells :These cells are large and have larger numbers of
nuclei with one to three small nucleoli and vacuolated cytoplasm.
 These cells are usually seen in small numbers mixed with other
inflammatory cells as part of a granulomatous reaction
 Common in many early focal inflammatory lesions in birds and reptiles
 Fibroblasts :frequently seen in association with inflammation.
 fibroblasts and fibrocytes are elongated cells with pointed tails of
cytoplasm. They have round or oval nuclei, indistinct nucleoli, and
moderate amounts of uniform, pale blue-staining cytoplasm. The
cytoplasmic boundaries are indistinct, giving a “wispy” appearance
 Reactive fibroblasts cannot be definitively distinguished cytologically from
low-grade spindle cell tumors.

40. Ancillary TechniquesAncillary Techniques
 FNAC samples for biomarker research isFNAC samples for biomarker research is
advancingadvancing
 microbiological, immunohistochemical,
radiological, biochemical and special staining
techniques
 electron microscopy, immunohistochemistry,electron microscopy, immunohistochemistry,
DNA ploidy, cytogenetics and FISHDNA ploidy, cytogenetics and FISH

41. Diagnostic Approach to General Cytology

42. Fluid Cytology
1.Centesis:-
 Centesis is the process of perforation with an aspirator,Centesis is the process of perforation with an aspirator,
trocar, or needle for abdominal, thoracic, synovial ortrocar, or needle for abdominal, thoracic, synovial or
cerebrospinal fluid collection (EDTA tube).cerebrospinal fluid collection (EDTA tube).
2.Catheterization:-
 transtracheal/ bronchial and prostatic washestranstracheal/ bronchial and prostatic washes

43. Peritoneal fluid cytological
examination
Indications
 AscitesAscites
 Inflammatory exudationInflammatory exudation
 Rupture of urinary bladderRupture of urinary bladder
 Colic and continuous weight loss in horsesColic and continuous weight loss in horses
(Thrall et al,1984)(Thrall et al,1984)

44. Cont….
Perform nucleated cell count :-Perform nucleated cell count :-
Normal peritoneal fluid has nucleated cells < 10000/microlit.Normal peritoneal fluid has nucleated cells < 10000/microlit.
 Bacteria suspected : stain with Gram’s stain and describeBacteria suspected : stain with Gram’s stain and describe
its morphology ( cocci, bacilli)its morphology ( cocci, bacilli)
 Congestive heart failure: TransudateCongestive heart failure: Transudate
Mono nuclear cell % increases.Mono nuclear cell % increases.
Macrophages
Neutrophil Cryptococcus org.

45. Pleural fluid examination
Indications
 NeoplasiaNeoplasia
 HemorrhageHemorrhage
 TransudationTransudation
 ChylothoraxChylothorax
Normally pleural fluid has nucleated cellsNormally pleural fluid has nucleated cells ≤≤ 10000/ micro lit.10000/ micro lit.
(Thrall et al,1984)(Thrall et al,1984)

46. BENIGN VS MALIGNANT
The presence of cells that are predominantly of the sameThe presence of cells that are predominantly of the same
population indicates normal tissue, hyperplasia, orpopulation indicates normal tissue, hyperplasia, or
neoplasianeoplasia
Benign neoplasia is characterized byBenign neoplasia is characterized by
 uniform population of cells from normal tissue.uniform population of cells from normal tissue.
 uniform cytoplasmic and nuclear size and shapeuniform cytoplasmic and nuclear size and shape
 uniform nuclear-cytoplasmic ratio.uniform nuclear-cytoplasmic ratio.
Benign tumor cell generally organized intoBenign tumor cell generally organized into
 Papillary structures,Papillary structures,
 Clusters SheetClusters Sheet
 Glandular formGlandular form
 Honey combingHoney combing
 Moulding patternMoulding pattern

47. Papillary Cluster
Honey Combing
Moulding

48. CYTOLOGIC CRITERIA OF
NEOPLASTIC CELLS
Diagnostic criteria of neoplastic cells will be based on :Diagnostic criteria of neoplastic cells will be based on :
 Cellular FeaturesCellular Features
 Nuclear FeaturesNuclear Features
 Cytoplasmic FeaturesCytoplasmic Features

49. Cellular Features of Malignant Cells
Hyper
chromasia
Anisocytosis Macrocytosis
Pleomorphism
High
N/C ratio
MALIGNANCY
CRITERIA

50. Anisocytosis Variation in cell size
Hypercellularity Increase cell exfoliation due to
decrease cell adherence
Pleomorphism Variable size and shape of the cell
Nuclear molding Deformation of nuclei by other
nuclei within same cell.
Cell polarity Lost
Cont…

51. Nuclear Features of Malignant Cells
Aniso
nucleoliosis
Nuclear
molding
Abnormal
Mitotic
figures
Macronuclei
Anisokaryosis
MALIGNANCY
CRITERIA

52. Hyperchromasia,
Anisokaryosis
High N/C ratio
Multinucleation
Atypical Mitosis
Nucleolar Features

53. Cytoplasmic Features of malignant cells
Keratinization
Intense
basophilia
Vacuolisation
Cannibalism
MALIGNANCY
CRITERIA

54. Vacuolisation Keratinization
Cannibalism

55.  Thyroid carcinomas -fairly uniform, well-differentiated cells,
in dogs tumors >3 cm are automatically considered malignant
 Similarly apocrine gland carcinomas, basal cell tumors,
melanomas, and proliferative lesions of the liver cannot be
distinguished b/w benign and malignant
 Lymphoma characterized by a uniform population of cells
that are larger than normal lymphoid cells. . If there is marked
variation and small lymphocytes are present, then that is more
typical of hyperplasia.
 Hepatoid glands- have more than one cell type (reserve and
terminal cells) and can therefore show variation in
morphology, and benign tumors of hepatoid glands can have a
similar mixture.
Exceptions to the rules of interpretation

56. Round cellRound cell Epithelial Epithelial MesenchymalMesenchymal
 Cellularity High High LowCellularity High High Low
 Cells Individual Clusters IndividualCells Individual Clusters Individual
 Shape Round Cuboidal SpindleShape Round Cuboidal Spindle
Types of tumors

57. Cytoplasmic
Vacuolation
Basal Cell
Carcinoma
Epithelial
Tumour
Round to
caudate
cells
Round & oval
Nucleolus
Adenoma,adenocacinoma
Anaplastic carcinoma
Sebaceous gland adenoma
Keratinized
Epithelial cells
PapillomaSquamous cell
carcinoma
Small Cuboidal
Cells+ Oval Nucleus
No
Round cells +
Single distinct
nucleus
Seminoma
Hepatoid
cells
Perianal gland
adenoma
Caudate cells + pale
cytoplasm
Ceruminous gland
adenocarcinoma
+
(Krithiga et al,2005)

58. Epithelial Tumors
 Aspirates of epithelial tumors tend to be of moderate to highAspirates of epithelial tumors tend to be of moderate to high
cellularity and contain large round to polygonal cells usuallycellularity and contain large round to polygonal cells usually
occurring in clusters or sheets.occurring in clusters or sheets.
 The cells have large prominent nuclei. Acinar or ductalThe cells have large prominent nuclei. Acinar or ductal
arrangements may be noted with adenomas andarrangements may be noted with adenomas and
adenocarcinomas.adenocarcinomas.
 Poorly differentiated epithelial neoplasms may producePoorly differentiated epithelial neoplasms may produce
more cells that occur individually as the cells lose theirmore cells that occur individually as the cells lose their
normal cellular attachmentsnormal cellular attachments

59. BenignBenign malignantmalignant
E.g. Basal cell tumorE.g. Basal cell tumor
Squamous cell tumor,Squamous cell tumor,
Sebaceous gland Adenoma ,Sebaceous gland Adenoma ,
Mammary & Prostate tumors,Mammary & Prostate tumors,
Transitional cell tumor.Transitional cell tumor.

60. Papilloma
Papilloma is finger like projectionPapilloma is finger like projection
*Cytology revealed keratinized epithelial cell with*Cytology revealed keratinized epithelial cell with
small round to oval nucleus.small round to oval nucleus.
*Nucleus was distinct*Nucleus was distinct
*Chromatin finely dispersed*Chromatin finely dispersed
*Cytoplasm was vacuolated. H & E stain*Cytoplasm was vacuolated. H & E stain

61.  They are the most commonly reported cutaneous tumorsThey are the most commonly reported cutaneous tumors
in dogin dog
 Have a site predilection for the skin of head, neck &Have a site predilection for the skin of head, neck &
shouldersshoulders.
 Basal cell tumors arise from multipotential germinalBasal cell tumors arise from multipotential germinal
epidermal cellsepidermal cells
 The tumors may be pigmented, and often contain cysticThe tumors may be pigmented, and often contain cystic
spaces.spaces.
 Basal cell tumors usually are benignBasal cell tumors usually are benign
Basal Cell Tumor

62. Cont.....
CYTOLOGYCYTOLOGY
*Clumps of tightly adherent cuboidal cells*Clumps of tightly adherent cuboidal cells
*Individual cell show cords or ribbons of small, uniform cell.*Individual cell show cords or ribbons of small, uniform cell.
*The cells have a high N:C ratio & mild anisokaryosis.*The cells have a high N:C ratio & mild anisokaryosis.
*Less cytoplasm*Less cytoplasm

63.  The neoplastic squamous cells exhibit Tadpole cells.The neoplastic squamous cells exhibit Tadpole cells.
* Anisocytosis , anisokaryosis and waxy blue cytoplasm* Anisocytosis , anisokaryosis and waxy blue cytoplasm
* Asynchrony of nuclear and cytoplasmic maturation,* Asynchrony of nuclear and cytoplasmic maturation,
* Occasional binucleate forms* Occasional binucleate forms
( Lumsden,2002)( Lumsden,2002)
Squamous cell carcinoma

64. Cytologically,Cytologically,
*Clusters of highly basophilic cells,*Clusters of highly basophilic cells,
* small centrally located nuclei and abundant foamy* small centrally located nuclei and abundant foamy
cytoplasmcytoplasm
* higher N:C ratio.* higher N:C ratio.
* cytoplasmic secretory material displace the nucleus to the* cytoplasmic secretory material displace the nucleus to the
periphery of the cellperiphery of the cell (signet ring cell)(signet ring cell)
Sebaceous Gland Tumor

65. Urinary neoplasm is transitional cellUrinary neoplasm is transitional cell
carcinoma.carcinoma.
*The cells are pleomorphic with*The cells are pleomorphic with
abundant cytoplasmabundant cytoplasm
*Anisocytosis & anisokaryosis.*Anisocytosis & anisokaryosis.
*the cytoplasm may contain small to*the cytoplasm may contain small to
large pale vacuoles.large pale vacuoles.
( Frost et. al, 2001 )
Transitional cell carcinoma

66. Cytology
• Epithelial cell clusters
• Nuclei spherical to
oval
• 1-2 nucleoli
• Foamy cytoplasm
ADENOMA

67. ADENOCARCINOMA - CYTOLOGY
 Cell clusters - high, round
to caudate, anisocytosis
 Spherical to oval nuclei
 Anisokaryosis & abnormal
shapes
 Multiple abnormal nucleoli
 Coarse chromatin
 Foamy cytoplasm

68. FIBROMA
•Presence of spindle cells
•Nuclei oval and elongated with indistinct
nucleolus
•Cytoplasm- tapering
(McCarthy et.al, 2003)

69.  Fine-needle aspirate from a mammaryFine-needle aspirate from a mammary
carcinoma in a dog.carcinoma in a dog.
 Anisocytosis and anisokaryosisAnisocytosis and anisokaryosis
are present (Wright stain).are present (Wright stain).
 Presence ofPresence of macro nucleolusmacro nucleolus
Mammary carcinoma

70. Hepatoid Gland Adenoma ,Perianal adenoma,
Circumanal gland adenoma
 Large, cuboidal epithelial cells look likeLarge, cuboidal epithelial cells look like liver cellsliver cells..
 Low N:C ratios, round nuclei, single distinct nucleolus,Low N:C ratios, round nuclei, single distinct nucleolus,
occassionaly binucleate.Mild anisocytosis andoccassionaly binucleate.Mild anisocytosis and
anisokaryosis presenceanisokaryosis presence
 Slightly granular blue cytoplasm.Slightly granular blue cytoplasm.

71. Fine-needle aspirates of the testicular masses.Fine-needle aspirates of the testicular masses.
 Highly cellular and vacuolated cells in sheets and palisades,Highly cellular and vacuolated cells in sheets and palisades,
Finely granular chromatin and prominent nucleoli,Finely granular chromatin and prominent nucleoli,
 Central round area of amorphous, deeply eosinophilic, hyalineCentral round area of amorphous, deeply eosinophilic, hyaline
material surrounded by a peripheral, rosette-like arrangementmaterial surrounded by a peripheral, rosette-like arrangement
of single or multiple rows of Sertoli cells. These structures wereof single or multiple rows of Sertoli cells. These structures were
suggestive ofsuggestive of Call-Exner bodiesCall-Exner bodies
Sertoli Cell tumour

72. Round Cell Tumor
 cells usually individual seen.
 circular cells with round nuclei, distinct cytoplasmic borders
 cells may be well differentiated.
Round

73. Yes Mast cell tumour
DISCRETE
ROUND
CELLS
Cytoplasmic
granules
No Cytoplasmic
vacuolations
Yes Transmissible
venereal tumour
No
Bean shaped and
indented nucleus
Multinucleated
giant cells
Yes Yes
Histiocytoma
a
b
dc
Histiocytic sarcoma

74. FNAC from the lesion on the lip of Dog.FNAC from the lesion on the lip of Dog.
 large round cells and variable numbers oflarge round cells and variable numbers of meta-chromatic
intracytoplasmic granules (purple) typical of neoplastic mast(purple) typical of neoplastic mast
cells.cells.
A small percentage of mast cell tumors, however, are composed almost entirely
of inflammation, hemorrhage, and edema with only a small number of mast
cells. These can be difficult to identify even by histology.
Mast cell tumor - dog

75.  Aspirate from an enlarged lymph node. TheAspirate from an enlarged lymph node. The
small blue droplets aresmall blue droplets are
““lymphoglandular bodies”lymphoglandular bodies” (arrows).(arrows).
• The nucleated lymphoblast , normal smallThe nucleated lymphoblast , normal small
Lymphocyte is indicated by red arrow.Lymphocyte is indicated by red arrow.
 Neoplastic lymphocytes withNeoplastic lymphocytes with convoluted nucleiconvoluted nuclei
typical of some cutaneous lymphomas.typical of some cutaneous lymphomas.
(Wright’s Giemsa stain, ×1000).(Wright’s Giemsa stain, ×1000).
Lymphoma

76. Plasmacytoma are tumors of plasmaPlasmacytoma are tumors of plasma
cells that occur outside the bonecells that occur outside the bone
marrow cavity.marrow cavity.
Cytologically,Cytologically,
Eccentric nucleus,Eccentric nucleus,
Abundant blue cytoplasm,Abundant blue cytoplasm,
Golgi zone look like osteoblasts,Golgi zone look like osteoblasts,
mostly are benign in nature.mostly are benign in nature.
Plasmacytoma/ multiple myeloma

77.  Fine needle aspirate from a mass on the penis of a dogFine needle aspirate from a mass on the penis of a dog
 Individual round cells with fairly abundant blue cytoplasm andIndividual round cells with fairly abundant blue cytoplasm and
uniform round nucleus with discrete cytoplasmic vacuoles.uniform round nucleus with discrete cytoplasmic vacuoles.
 Key to the diagnosis is: cytoplasmic vacuoles and Location-
genital area.
Transmissible venereal tumor

78. “histiocytoma" is a benign neoplasm of monocytic cells.histiocytoma" is a benign neoplasm of monocytic cells.
Tends to be ulceratedTends to be ulcerated
cutaneous histiocytomas are derived from a more specializedcutaneous histiocytomas are derived from a more specialized
population of cells called Langerhans cells.population of cells called Langerhans cells.
Histiocytes are mainly mature macrophages that reside inHistiocytes are mainly mature macrophages that reside in
connective tissue.connective tissue.
Cutaneous histiocytoma

79.  In histiocytoma ,In histiocytoma ,
*Cells with a round to oval centrally located nucleus*Cells with a round to oval centrally located nucleus
*Moderately abundant blue cytoplasm that lacks vacuoles*Moderately abundant blue cytoplasm that lacks vacuoles
and granules.and granules.
*A binucleated cell and mitotic figure are present*A binucleated cell and mitotic figure are present
(Wright stain)(Wright stain)

80. Mesenchymal tumors
Mesenchyma is the meshwork of embryonal tissue that formsMesenchyma is the meshwork of embryonal tissue that forms
the body C.T. and vessels.the body C.T. and vessels.
Mesenchymal tumors exfoliate poorly because the cells areMesenchymal tumors exfoliate poorly because the cells are
embedded in extracellular matrix such as fibrous connectiveembedded in extracellular matrix such as fibrous connective
tissue, cartilage, or bone.tissue, cartilage, or bone.
Malignancy of mesenchymal origin are called sarcoma.Malignancy of mesenchymal origin are called sarcoma.

81.  Cytologically,Cytologically,
cells are predominantlycells are predominantly fusiformfusiform
have elongated nuclei andhave elongated nuclei and
cytoplasmic tailscytoplasmic tails (spindle cells).(spindle cells).
 so its either a mesenchymalso its either a mesenchymal
(spindle cell) tumor or granulation(spindle cell) tumor or granulation
tissuetissue
Cont…
Mesenchymal lesions should be submitted for HistopathologyMesenchymal lesions should be submitted for Histopathology
active fibroblasts are difficult to differentiate from neoplasticactive fibroblasts are difficult to differentiate from neoplastic
fibroblasts and may even have features that suggestfibroblasts and may even have features that suggest
malignancy.malignancy.

82.  Lipomas are benign tumors but they can occasionallyLipomas are benign tumors but they can occasionally
infiltrate between muscle masses.infiltrate between muscle masses.
 Fine-needle aspirate of a canine Lipoma, lipocytes have aFine-needle aspirate of a canine Lipoma, lipocytes have a
balloon-like appearanceballoon-like appearance with voluminous cytoplasm and awith voluminous cytoplasm and a
small peripherally displaced nucleussmall peripherally displaced nucleus
Lipoma

83. Cont….
 Liposarcoma are rare malignant tumors of white adiposeLiposarcoma are rare malignant tumors of white adipose
cell lipoblastscell lipoblasts
Malignant lipocytes appear round to polyhedral withMalignant lipocytes appear round to polyhedral with
vacuolated basophilic cytoplasm,vacuolated basophilic cytoplasm,
A round nucleus (anisokaryosis), and occasional prominentA round nucleus (anisokaryosis), and occasional prominent
nucleoli and mitotic figure is present.nucleoli and mitotic figure is present.

84.  Dermal melanomas are common in dogs and horses.Dermal melanomas are common in dogs and horses.
 The tumor aspirate contains predominantly individualThe tumor aspirate contains predominantly individual
Melanocyte (round, oval, stellate or spindle shaped).Melanocyte (round, oval, stellate or spindle shaped).
 Tumor cells with giant nuclei are often seen. MelaninTumor cells with giant nuclei are often seen. Melanin
pigment appears aspigment appears as brown-black to green-blackbrown-black to green-black
cytoplasmic granulescytoplasmic granules of irregular size and shape.of irregular size and shape.
Melanoma

85.  hemangiosarcomas are tumors of blood vessel endothelium andhemangiosarcomas are tumors of blood vessel endothelium and
are contiguous with the blood vascular systemare contiguous with the blood vascular system
 Neoplastic endothelial cells aspirated are large spindle toNeoplastic endothelial cells aspirated are large spindle to
polyhedral cells. The nucleus is round to oval and contains one orpolyhedral cells. The nucleus is round to oval and contains one or
more prominent nucleoli. The cytoplasm appears dark blue andmore prominent nucleoli. The cytoplasm appears dark blue and
usually contains many small, discrete, non staining vacuolesusually contains many small, discrete, non staining vacuoles
Hemangiosarcoma

86.  Osteosarcoma (OSA) is a rapidly growing, destructive neoplasmOsteosarcoma (OSA) is a rapidly growing, destructive neoplasm
of boneof bone
 Fine-needle aspirates of OSA contain mesenchymal cells thatFine-needle aspirates of OSA contain mesenchymal cells that
appear round, plump, or fusiformappear round, plump, or fusiform
 Individual neoplastic cells may display anisocytosis,Individual neoplastic cells may display anisocytosis,
anisokaryosis, karyomegaly, eccentrically located nuclei, largeanisokaryosis, karyomegaly, eccentrically located nuclei, large
nucleoli, and basophilic vacuolated cytoplasm .nucleoli, and basophilic vacuolated cytoplasm .
Osteoclast
Osteoid matrix
Osteosarcoma

87.  Chondrosarcoma are the second most common tumor ofChondrosarcoma are the second most common tumor of
bone and are difficult to differentiate from osteosarcomasbone and are difficult to differentiate from osteosarcomas
cytologicallycytologically
 Individual neoplastic chondroblasts have cytologic featuresIndividual neoplastic chondroblasts have cytologic features
similar to those of osteosarcoma cells.similar to those of osteosarcoma cells.
Chondrosarcoma

88. Based on the type of cell in smear following conclusions canBased on the type of cell in smear following conclusions can
be drawn.be drawn.
 Lymph node enlarged : Hyper plastic lymph nodeLymph node enlarged : Hyper plastic lymph node
 Lymph node enlarged with : Reactive lymph nodeLymph node enlarged with : Reactive lymph node
increase in Lymphoblast, Mincrease in Lymphoblast, MØØ,,
with haemosiderin and debriswith haemosiderin and debris
 Non lymphoid cells with : Metastatic NeoplasiaNon lymphoid cells with : Metastatic Neoplasia
malignant cellsmalignant cells
 Majority of cells are : Lymphoid NeoplasiaMajority of cells are : Lymphoid Neoplasia
lymphoblastslymphoblasts
(Roshni(Roshni et alet al, 2007), 2007)

90. Lymph Node Cytology
Aspirate from a hyperplastic lymph node. Numerous smallAspirate from a hyperplastic lymph node. Numerous small
lymphocytes (arrows) are present. A lymphoblast (double arrows)lymphocytes (arrows) are present. A lymphoblast (double arrows)
and prolymphocyte (arrowhead) are present also. Numerousand prolymphocyte (arrowhead) are present also. Numerous
lymphoglandular bodies are present in the background of thelymphoglandular bodies are present in the background of the
smear.smear.

91. Metastatic Neoplasia
Cytology smear showing clusters of keratinizing squamousCytology smear showing clusters of keratinizing squamous
carcinoma indicating metastasis in the lymph node.carcinoma indicating metastasis in the lymph node.

92. lymph node aspiration show majority of large lymphoblast, withlymph node aspiration show majority of large lymphoblast, with
one to several prominent nucleoli (Wright's-Giemsa stain)one to several prominent nucleoli (Wright's-Giemsa stain)
lymphoblastic lymphoma

93. Hyperplasia
Reactive changes
Repair

94. FNAC immunocytochemistryFNAC immunocytochemistry
estrogen receptorsestrogen receptors

95. FISH: amplification of HER2 gene (green spots).

96. Advantages of Cytology
 Samples collected easily and quickly prepared stained andSamples collected easily and quickly prepared stained and
interpreted. less expensive and little or no risk to the patientinterpreted. less expensive and little or no risk to the patient
 Cytologic examinations early identify disease process, neoplasiaCytologic examinations early identify disease process, neoplasia
vs. inflammation.vs. inflammation.
 This may allow the treatment of the tumor when it is stillThis may allow the treatment of the tumor when it is still
very small, greatly increasing the chances of a cure.very small, greatly increasing the chances of a cure.

97. Limitations of Cytology
 The most significant disadvantage of cytology is the absence ofThe most significant disadvantage of cytology is the absence of
tissue architecture.The arrangement of neoplastic cells withintissue architecture.The arrangement of neoplastic cells within
tissues is critical in determining the diagnosis of many types oftissues is critical in determining the diagnosis of many types of
tumors.tumors.
(M.L. Wellman,1990)(M.L. Wellman,1990)
 It is not always possible to:It is not always possible to:
localize neoplastic lesion,distinguishe invasive cancer,localize neoplastic lesion,distinguishe invasive cancer,
distinguishe reactive dysplastic and neoplastic changes,distinguishe reactive dysplastic and neoplastic changes,
determine tumor type.determine tumor type.
(Sr. Kamenica 2002)(Sr. Kamenica 2002)

98. complicationscomplications
 hematomashematomas
 AbscessesAbscesses
 PneumothoraxPneumothorax
 FistulasFistulas
 pain, edema, psychological trauma, smallpain, edema, psychological trauma, small
 hemorrhages and vasovagal reactionshemorrhages and vasovagal reactions

99. CONCLUSION
FNAC is a simple, rapid and in-expensive aid which
can be used in all clinics and hospitals
Impression smears can be made from incisional and
excisional biopsies for cytological examination
Haematoxylin and eosin staining gives familiar
tinctorial properties as that of histopathology
Romanowsky stains used on air-dried smears gives
good nuclear and nucleolar details
Expertise can be gained with experience by
following a systematic procedure

100. Vaginal cytologyVaginal cytology

104. ReferencesReferences
 Damadian et al.Damadian et al. “Method of Conducting a Needle Biopsy Procedure”.“Method of Conducting a Needle Biopsy Procedure”. United States PatentUnited States Patent
No. US 7,008,383 B1: March 7, 2006.No. US 7,008,383 B1: March 7, 2006.
 ““Fine Needle Aspiration”.Fine Needle Aspiration”. Medax Medical Devices: 2005. http://www.medaxmedical.comMedax Medical Devices: 2005. http://www.medaxmedical.com
 ““Fine Needle Aspiration”.Fine Needle Aspiration”. American Academy of Otolaryngology: 2006.American Academy of Otolaryngology: 2006.
http://www.entnet.orghttp://www.entnet.org
 ““Fine Needle Aspiration Biopsy (FNA)”.Fine Needle Aspiration Biopsy (FNA)”. Virtual Cancer Centre: 2006.Virtual Cancer Centre: 2006.
http://www.virtualcancercentre.comhttp://www.virtualcancercentre.com
 Hopper MD, K. et al.Hopper MD, K. et al. “Fine-Needle Aspiration Biopsy for Cytopathologic Analysis: Utility of“Fine-Needle Aspiration Biopsy for Cytopathologic Analysis: Utility of
Syringe Handles, Automated Guns, and the Nonsuction Method”.Syringe Handles, Automated Guns, and the Nonsuction Method”. Radiology: December 1992,Radiology: December 1992,
(819-824).(819-824).
 Kelcz MD, Frederick. Department of Radiology.Kelcz MD, Frederick. Department of Radiology.
 Mladinich DVM, MS, C. et al.Mladinich DVM, MS, C. et al. “Evaluation and Comparison of Automated Biopsy Devices”.“Evaluation and Comparison of Automated Biopsy Devices”.
Radiology: September 1992, (845-847).Radiology: September 1992, (845-847).
 Suen, K. “Fine-Needle Aspiration Biopsy of the Thyroid”. Canadian Medical AssociationSuen, K. “Fine-Needle Aspiration Biopsy of the Thyroid”. Canadian Medical Association
Journal: September 2002, (491-495).Journal: September 2002, (491-495).

105. Cytology is not a substitute
for histology
but……….
Thin needle aspiration biopsy is a procedure
whose time has come” and the pathologists not
already versed in the technique will come under
increasing and compelling pressure to provide it