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Cytological Techniques Flashcards

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1
Q

– refers to diagnostic techniques that are used to examine cells from various body sites to determine the cause or nature of disease

A

Cytopathology

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2
Q

19th century

A

The First Era

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3
Q

The Second Era: __ and __
___ - Father of cytopathology

A

development and expansion
- Dr George Papanicolaou

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4
Q

The Third Era : __
- ___

A

consolidation
- Dr Leopold Koss Diagnostic

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5
Q

The Fourth Era

A

The Bethesda System for Reporting Cervical/Vaginal Cytology Diagnoses

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6
Q

diagnostic cytology
– microscopic examination of cells from different body sites for ___ .
– includes _ and _

A

diagnostic purposes
- exfoliative cytology, and fine needle aspiration (FNA)

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7
Q

– microscopic study of cells that have been desquamated from epithelial surfaces

A

Exfoliative cytology

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8
Q

EC - Recommended
(5)

A

• Detection of malignant cells
• Detection of precancerous cervical lesions in women
• Assessment of female hormonal status
• For determination of genetic sex
• For detection of infectious agents

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9
Q

Gynecologic

A

• Cervicovaginalsmear (Pap smear)
– vaginal, endometrial and endocervical smears

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10
Q

Non-gynecologic

A

• Prostatic and breast secretions
• Gastric or bronchial secretions
• Pleural and peritonial fluids
• Sputum
• Smears of urine sediments
• CSF specimen

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11
Q

Collection and Preparation of

Specimen Specimen for examination may be taken from various body regions

A

Gynecologic
Non- Gynecologic

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12
Q

Methods of Smear Preparations

A

ü Streaking-
ü Spreading-
ü Pull apart -
ü Touch/ Impression/ Imprint/Abraded smear

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13
Q

Smearing techniques
• Characteristics the ideal smear:

A

– should be evenly spread
– uniformly thin and flat
> to enable rapid drying and fixation
>to permit optimal penetration of stain

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14
Q

Adhesive Agents for Cytologic Method

A

– Pooled Human serum or Plasma
– Celloidin Ether Alcohol
– Leuconostoc Culture

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15
Q

-
-

A

– Urinary sediment
– Bronchial lavage specimen
– Specimen that utilizes proteolytic enzymes

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16
Q

Prepared smears must be immediately fixed
• ___ -for all types of effusion if there is some delayed in smearing
• Saccomano preservative (_ and_ )

A

Fixation
- 50%alcohol
- 50% alcohol and 2% carbowax

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17
Q

• Common Fixatives

A

– Ether- alcohol - best fixatives
– 95% ethanol - routine preparation

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18
Q

– selectively lyses __
– Used in heavily blood-stained specimens.

A

Carnoy’s fluid
- erythrocytes

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19
Q

Otherfixatives

A

– glutaraldehyde or formalin
– Polyethylene glycol in alcohol

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20
Q

• provides a protective waxy coating for postal despatch

A

– Polyethylene glycol in alcohol

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21
Q

• This method uses__ and __
• The cells are isolated via a series of centrifugation steps to concentrate the cells into a small suspension.

A

Cell concentration techniques
-cytospin and sedimentation preparations

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22
Q

• __ is a paraffin-embedded specimen derived from different fluids and aspirated materials.

A

Cell Block Technique
- Cell block

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23
Q

• Used for establishing a more definitive cytopathologic diagnosis.
– “__”
• Serve as a bridge between cytology and histology

A

cell block
- microbiopsy

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24
Q

Uses of cell block (5)

A

• Architectural evaluation
– (histologic pattern of tumor)
• Categorization of tumors that are not possible for smear
• Special stains and Immunohistochemistry
• Immunophenotyping, molecular studies
• As archival material for future studies

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25
Romanowsky type stains (for air dried slides)
– Wright’s Giemsa stain – May Grunwald – Giemsa stain – Diff- Quick
25
– (for immediate fixated slides)
Papanicolaou stains
26
Stains
Romanowsky type stains Papanicolaou stains
27
Staining Methods in Cytology • Papanicolaou staining method • It is a polychrome staining reaction that results in __,___,__
- well stained nuclear chromatin, differential cytoplasmic counterstaining and cytoplasmic transparency
28
• Cytoplasmic stains
– OG-6 (monochrome stain) – EA-36 (polychrome stain)
29
• Formalin-fixed cytology preparations must be stained with either _ or _
H&E or Papanicolaou stain
30
__ = nuclear stain  __ - cytoplasmic stain for MATURE (superficial cells)  __ = for IMMATURE vaginal cells > (_, _)
Hematoxylin OG 6 EA 36 or 50 > parabasal, intermediate cells
31
• Respiratory Tract specimen – Principle __
These specimen are obtained to exclude the possibility of malignancy or infectious agents
31
Stains for cytochemistry (5)
1 Periodic Acid Schiff (PAS) for glycogen 2 Perl’s stain for hemosiderin 3 Alcian Blue for mucins 4 Grocott Methenamine Silver (GMS) for fungal organisms 5 Ziehl-Neelsen stain for acid fast bacilli.
32
– Type of Specimen of Respiratory Tract specimen
• sputum, Bronchoalveolar Lavage, BW, BB
33
– Cytologic Collection and Preparation • Sputum collection - -
– at least 3 consecutive morning – Collectt every morning in a wide-mounted jar containing Saccomano fluid
33
Gastrointestinal specimen • Principle
The collection is usually done to exclude the possibilty of malignant tumors
34
• Types of specimen of Gastrointestinal specimen
GL, GB, FNA
35
Cytologic Collection and Preparation of Gastrointestinal specimen
8 hours fasting before the gastric washing
36
• The presence of malignant cells in serous effusions usually indicate __
Peritoneal, Pleural and Pericardial Fluids - metastatic involvement
37
• Jelly-like clots forming after removal may prevented by adding __
Peritoneal, Pleural and Pericardial Fluids -300 units of heparin for every 100 ml of aspirates
38
• diagnosis of malignancy, in urothelial origin • __ are rarely found in urinary specimens • Types of specimen 1 2 3
Urinary Tract Specimen - Prostatic carcinomas -Voided urine – Catheterized specimen – Washing from bladder or renal pelvis
39
• important diagnostic value in patientswith a known history of cancer • __ is the first presentation of cancer of unknown origin
Body Cavity Effusions - Positive effusion of malignancy
39
- it has an extremely low diagnostic yield for diagnosis of __. - __ is usually a result of hormonal imbalance in young patients, and when the secretion is bloody a ___ should be considered clinically. - Specimen should be smeared on a clean glass slide, and immediately placed in fixative
Smears of Breast Secretion - breast carcinoma - Spontaneous nipple discharge - benign intraductal papilloma
40
Types of Specimen of Body Cavity Effusions
- Pleural fluid - ascitic/abdominal fluid - Peritoneal washing - Pericard - Cerebrospinal fluid
41
Cytologic Collection & Preparation - Specimen is collected in a __ - Specimen place in a __ if there is adelayed - _ does not interfere with cytologic preparation - _ IS necessary for cytologic evaluation
- clean- non-sterile dry container - refrigerator - Heparin - 1ml of CSF
42
is the study of cellular samples obtained from organs that do not shed cells spontaneously, such as (8) - It is useful in lesions that are easily palpable, like (6)
Fine Needle Aspiration (FNA) - breast, thyroid, lymph nodes, liver, lungs, skin, soft tissues and bones - growth of skin, subcutaneous soft tissue tumors, thyroid, lymph nodes, salivary glands and breast
43
Slide Preparation recommend preparation of maximum 4 slides: using 1-2 drops on each slide and using slide-pull technique (similar to the technique used for peripheral blood smears).
Fine Needle Aspiration (FNA)
44
Slide Fixation Immediate fixation is important
Fine Needle Aspiration (FNA) 95% alcohol spray fixative alcohol-acetone (Papstain) Formalin
45
is the study of cellular samples obtained from organs that do not shed cells spontaneously, such as (8) - It is useful in lesions that are easily palpable, like (6)
Fine Needle Aspiration (FNA) - breast, thyroid, lymph nodes, liver, lungs, skin, soft tissues and bones - growth of skin, subcutaneous soft tissue tumors, thyroid, lymph nodes, salivary glands and breast
46
Cytologic Collection and Preparation (6)
– For conventional Pap smears – Endocervical brush – samples of endocervical canal – Vaginal scrape – for patient with hysterectomy – Lateral VS – hormonal evaluation – Four quadrant VS – localization of vaginal adenosis – Vulvar scrape – detection of herpetic lesions of carcinoma
47
Gynecologic specimen • ___ -junction of endocervical/ectocervical mucosa – Detection of __ and __ • Type of specimen – _,_,_
T-Zone(TransformationZone) - dysplasias and carcinomas - squamous, columnar and metaplastic cells
48
• Transparent blue staining of cytoplasm is obtained • Excellent nuclear detail • Color range is predictable and of great valuein identification, and cell classification • Valuable in comparing cellular appearances in smears
Staining Method- Papanicolau Method
49
Cells found in Cervico – __
vaginal smears
50
– Polygonal squamous cells with __ in diameter – __ -staining cytoplasm – dark pyknotic nuclei, less than _ in diameter – true acidophilia • under _
Cells found in Cervico-vaginal smear • Mature superficial cell - 45-50 um - pale, pink - 6u - estrogen influence
51
false acidophilia observed due to
Pseudoacidophilia - drying of smears esp before fixation - prolapse and drying of vaginal epithelium - infection - chemicals
52
– Round,oval or boat shaped cells – With __ – Nucleus pushed aside or towards cellmembrane – With double-walled boundary appearance (___ at the periphery)
Pregnancy cells - translucent basophilic cytoplasm - deep blue stain of the cytoplasm
52
• Medium polyhedral or elongated cells with basophilic vacuolated cytoplasm
Intermediate cell
53
– Boat-shaped intermediate cells with strong tendency to fold or curl on edges – suggest of the combined _ – Seen in __,__,__
Navicular cells - estrogen-progesterone effect - latter half of menstrual cycle, during pregnancy, and menopause
54
• Round to oval cells with small dense basophilic cytoplasm • __ in diameter, smaller than IC, have a larger __ • Found from __ of age to __ • found in _,_,_
Parabasal cells - 15-30 um - vesicular nucleus - two weeks-> puberty - After childbirth, with abortions and after menopause
55
• Similar to fried fresh eggs with sunny side up
Parabasal cells
56
• occur in large groups or small sheets • cytoplasm stained __ finely vacuolated • present a __ apperance
Endocervical glandular cells - pale blue/gray - honeycomb
56
• Small cells, slightly cylindrical with less basophilic cytoplasm • tightly packed groups of 3 or more • found during and __ • shed in response to __
Endometrial cells - 1-10 days after menstruation - ovarian hormone
57
- yeast - __ commonly seen in _,_,_,_,_
Candida albicans - candidiasis - diabetic patients, pt’s taking oral contraceptives, immunocompromised states, leukemia and lymphoma
58
Pear-shaped
Trichomonas vaginalis
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• G(+), slender rod-shaped m.o. • comprising the most commont organism found in a normal vaginal flora • staining_ to _ with Pap’s stain • numerous in the __ a during pregnancy
Doderlein Bacillus (Lactobacillus acidophilus) - blue to lavender - Corpus luteum phase
60
• Mucus on drying,exhibit a “fern”or palm-leaf pattern (__)due to ___ under the influence of _.
Ferning - arborization - formation of salt crystals in high NaCl concentration in cervical mucus - estrogen
61
Quantitation Evaluation for Vaginal Cytology
• Acidophilic Index • Pyknotic index • Maturation index
62
– is the percentage of cells staining pink-orange to red with Pap’s smear – not reliable index due to the possible __
• Acidophilic Index - pseudoacidophilia
63
– is the percentage of cells with shrunken, dark, small structureless nuclei
• Pyknotic index
64
– is the percentage of cells from the main layers of vaginal epithelium: _,_,_ >_ is used as criterion for mature superficial cells
• Maturation index > superfacial, intermediate and deep (parabasal and basal) cells - Pyknosis
65
CLASSIFICATION OF PAP TEST RESULTS
ClassI Negative for Malignant cells Class II Atypical cells present, butt Negative forMalignancy ClassI lII Suspicious for Malignant Cells Class IV Strongly Suggestive for Malignant Cells Class V Conclusive for malignant Cells
66
– is the percentage of cells from the main layers of vaginal epithelium: _,_,_ >_ is used as criterion for mature superficial cells
• Maturation index > superfacial, intermediate and deep (parabasal and basal) cells - Pyknosis
67
T h e BETHESDA SYSTEM Specimen Adequacy:_,_,_
Satisfactory Limited Unsatisfactory
68
Specimens to which the following conditions apply will be rejected: (6)
1 .Specimen is submitted without a requisition. 2 .Specimen is not labeled with the patient name. 3 .The patient name (or other identifying information) on the specimen and requisition do not correspond. 4 .The specimen i s labeled appropriately but th e requisition is no t labeled. 5 .The specimen slide(s)i s(are) irreparably broken. 6 .Specimen is submitted from an unauthorized source
69
the bethesda system General Categorization:
Negative for Intraepithelial lesion or malignant cell Epithelial cell abnormality
70
Descriptive Diagnosis of bethesda system
Atypical squamous cells of unknown significance Low grade squamous intraepithelial lesion High gr a de s q u a m o us intraepithelial lesion Squamous Cell Carcinoma Glandular cell abnormality Atypical glandular cells Adenocarcinoma Others
71
Papsmear specimens are considered satisfactory for interpretation if there are:
•Adequate numbers of well-visualized squamous cells present •Adequate numbers of well-visualized endocervical cells or squamous metaplastic cells (from the transformation zone). •Less than 50% of the cells obscured by blood ori nflammation •Properly labeled specimens
72
Papsmear specimens are considered unsatisfactory for interpretation if there are
• Inadequate numbers o f well-visualized squamous cells present • Inadequate numbers of well-visualized endocervical cells or squamous metaplastic cells (from the transformation zone). •More than 75% of the cells obscured by blood or inflammation •Improperly labeled specimens Usually,these smears are recommended for repeat sampling.

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