CYTOLOGIC TECHNIQUES Flashcards
1
Q
simply means microscopic examination of cells from different body sites for diagnostic purposes.
A
Diagnostic Cytology
2
Q
includes exfoliative cytology and fine needle aspiration.
A
Diagnostic cytology
3
Q
For _______to be of diagnostic value, the sample must include representative material and the method used to process the specimen must give rapid results, without sacrificing cellular detail.
A
cytologic examination
4
Q
_____ and _____are most important in cytological interpretation.
Cytologists rely heavily on the quality and appearance of the____.
A
Consistency and reliability
stain
5
Q
Specimen for cytologic examination may be derived from various sources:
A
- Exfoliative cytology
- Fine needle aspiration
- Body fluids
6
Q
deals with the microscopic study of cells that have been desquamated from epithelial surfaces.
A
Exfoliative cytology
7
Q
may be found in smears that have been spontaneously shed or physically removed from epithelial and mucous membranes.
A
Exfoliated cells
8
Q
The cytological specimens collected from female genital tract include(3)
A
cervical smear
vaginal smear
endometrial smear
9
Q
is observed in normal cells due to constant growth and replacement with new cells - more readily observed in malignant tumor cells.
A
Spontaneous exfoliation
10
Q
Specimen can be collected from the epithelial surfaces by (3)
A
lightly scraping the surface
by swabbing
aspirating or washing the surfaces
11
Q
Exfoliative cytology
• Assessment of female hormonal status in case of sterility and endocrine disorders.
This is achieved by microscopic evaluation for determination of________, based on examination of smears taken from the______
A
maturation index (MI)
lateral vaginal walls
12
Q
Exfoliative cytology
• Detection of malignant cells in body fluids, mainly used for_____
A
staging cancer
13
Q
Exfoliative cytology
• Detection of precancerous cervical lesions in women using..
A
cervicovaginal smear/Pap smear
14
Q
Exfoliative cytology
• For determination of genetic sex -most of the nuclei of females exhibit conglomeration of chromatin, representing XX chromosomes (Barr body), which may be demonstrated in the smears from….
A
buccal or vaginal mucosa
15
Q
T or F
Exfoliative cytology can be used for detection of infectious agents.
A
True
16
Q
Smear Preparation
Smears should be made from___ material, prepared in the doctor’s office or radiology units.
A
fresh
17
Q
Histotechnicians sometimes perform special stains on cytology smears, blood films and cytopreps from other departments within the laboratory.
Increasingly, the commonly received cytoprep is that of the “______”
A
thin prep.
18
Q
“thin prep”
These smears are wet-fixed in______ immediately after preparation to preserve the fine structure of the chromatin and help in the evaluation of nuclear changes.
A
95% ethanol
19
Q
is avoided with smears for cytological detection of neoplasia because it changes the appearances of the cells.
A
Air drying
20
Q
Slides bearing blood or bone marrow smears, on the other hand, are usually…
A
air-dried
21
Q
Cancer of the_____ is the commonest cancer that can be detected even at the pre-invasive stage.
A
uterine cervix
22
Q
involves the microscopic examination of cell samples that have been taken primarily from the ecto- and endocervix, smeared on glass slides and stained by the Papanicolaou (Pap) method.
A
Cervical cytology screening
23
Q
True or False
Cervical cytology is only for screeninh
A
True
24
Q
The patient should abstain from coitus, not douche the vagina for at least_____, and not apply intravaginal preparations for at least ____before the examination.
A
a day
one week
25
The use of______ for collection of cervical smear is to be discouraged because of the drying artifacts and loss of cells that are caused by this method.
cotton swab
26
is preferable to plastic spatula, because of its mildly rough surface that can collect more material.
Wooden spatula
27
is used strictly for taking materials from endocervix.
Endo-cervical brushing
28
are often used for ulcerated surface lesions to allow an immediate assessment of the lesion before fixation and processing of the tissue
sample.
The preparations should be quickly air dried and then stained. It is also indicated in the case of tumors specially of lymph nodes.
Impression smears
29
Sputum Smear
• Obtain at least________ specimens.
• Collect early morning sputum by a deep cough in a______ jar containing______
3 consecutive morning sputum
wide -mouthed
Saccomanno fluid
30
Saccomanno fluid
50% ethyl alcohol
2% carbowax
31
If the patient cannot cough up sputum spontaneously, induced sputum should be collected using inhalation of an aerosol solution for_____ to produce a deep cough sample.
20 minutes
32
Sputum smear
If a more extensive study is to be made, it is recommended that a minimum of ____slides be prepared, and one is air dried for_____
At least two slides should be stained by______ method.
2-4; Giemsa staining
Papanicolaou method
33
Sputum smear
With the end of a wooden applicator stick, these samples are evenly spread on the slides and immediately placed in fixative for a minimum of…
one hour
34
Sputum collected should have been coughed from "______" to ensure that the specimen is a true representative of the sputum and not just saliva.
The mucus present preserves the cell constituents; however, it is highly recommended that the specimen be as fresh as possible when examined, if better cellular detail is to be appreciated.
down-deep
35
________are found on the sputum smears from a deep cough.
Absence of these suggests "saliva" rather than sputum and results an unsatisfactory specimen.
Alveolar macrophages
36
Bronchial Brushing specimen is directly smeared onto two labeled slides by______ (usually done by a bronchoscopic specialist at the bronchoscopy laboratory).
pull technique
37
________specimens are freshly collected in the bronchoscopy collection container and hand delivered to the laboratory.
Bronchial Washing
38
Bronchoscopy specimen
Care should be taken to collect specimen taken directly from the bronchus, to insure that the smear contains_________ from the bronchial mucosa, not just red blood cells and leukocytes.
epithelial cells (ciliated bronchial cells)
39
are quite difficult to make due to inaccessibility of the specimen and the presence of gastric juices which have a deleterious effect on the morphology of exfoliated cells.
Gastric smear preparations
40
Gastric smear preparations
Smears are usually collected by ______ and ______
The collected specimen is then sent to the laboratory. Specimen should be examined as soon as possible since any delay of more than_____, before fixation, will digest the cells and make the specimen unsatisfactory for evaluation.
simple irrigation and aspiration technique.
I /2 hour
41
Gastric smear prep
The patient should have fasted for at least______ before gastric washing is performed. Esophageal washings are to be examined immediately.
8 hours
42
T or F
Cytologic examination of nipple discharge has an extremely high diagnostic yield for diagnosis of breast carcinoma.
False
Low
43
Spontaneous nipple discharge is usually a result of hormonal imbalance in young patients, and when the secretion is bloody a_______ should be considered clinically.
The spontaneous nipple discharge should be smeared on a clean glass slide, and immediately placed in fixative.
benign intraductal papilloma
44
In women, except during lactation and the immediate post-lactation period, any discharge from the nipple is abnormal .
The nipple discharge is usually due to a benign breast lesion such as_____and _____, or due to endocrine problems.
duct ectasia and papilloma
45
However the major value of cytologic examination of nipple discharge is potential detection of_______ in a patient with clinically undetected
carcinoma.
malignant cells
46
Breast secretion
Collection Technique:
Gently strip the subareolar area and nipple using the thumb and forefinger.
Place the labeled slide upon the nipple and draw it quickly across the nipple. If more than a drop is collected, use another slide to smear with a _____technique.
Immediately drop slide in a bottle of_____ or _____
pull-up
95% isopropanol or use spray fixative.
47
Breast secretion collection technique
If the secretion is scanty in amount, smears should be restricted to a small area of the slide to prevent____\.
Secretions obtained from both breasts should be properly identified as left or right.
For localized breast lesions producing discharge upon pressure, the secretion may be smeared directly on the slide after expressing material from the breast.
drying
48
is the study of cellular samples obtained from organs that do not shed cells spontaneously, such as breast, thyroid, lymph nodes, liver, lungs, skin, soft tissues and bones.
Fine needle aspiration
49
It is useful in lesions that are easily palpable, like growth of skin, subcutaneous soft tissue tumors, thyroid, lymph nodes, salivary glands and breast.
Fine needle aspiration
50
FNA
The basic technique uses a_____ gauge and ____ ml. The needle is inserted into the lesion and repeatedly redirected to sample a number of areas while applying a small amount of suction on the syringe.
25-gauge needle and a 10-ml syringe
51
FNA
Tissues composed of ________tightly adhere to each other and do not exfoliate easily, requiring a larger bore needle and increased suction may be necessary.
The sample should be air dried as quickly as possible to reduce the effects of shrinkage.
mesenchymal cells (connective tissue)
52
Deeply seated lesions such as lung, mediastinum, abdominal organs (liver, pancreas, etc.) and retroperitoneal organs (kidney, adrenal, lymph nodes) are performed under…
laparoscopy computerized tomography (CT scan) ultrasound (sonography).
53
FNA
When a solid lesion is aspirated, usually a few drops from the____ of the needle has the most diagnostic material for cytologic evaluation.
tip
54
FNA
On the other hand when the specimen is______, the diagnostic cells are diluted and hard to find on a direct smear.
We recommend preparation of maximum____ slides: using
1-2 drops on each slide and using slide-pull technique (similar to the technique used for peripheral blood smears).
Then rinse the needle in a preservative solution such as______, and send it to the laboratory. It will be processed like any other body effusion specimen at the cytology laboratory.
bloody
4 slides
Saccomano fluid
55
An ideal aspirate is of_____ consistency with numerous cells suspended in a small amount of tissue fluid without admixture with blood.
In lymph node aspiration, a cell suspension can be prepared in addition to direct smears.
creamy
56
• Colloid, mucin and smears to be stained with_________should air dried. Smears that are not correctly made and dried quickly will produce artefacts.
Rapid stains like_______, are particularly useful in preliminary assessment of adequacy of the sample before the patient is released.
hematological stains (such as May-Grunwald-Giemsa)
Diff Quik (2-3 minutes)
57
• Smears to be stained by ______ or ______should be rapidly fixed in alcohol (wet fixation) to show nuclear details and allow better identification of malignant cells.
If the smears are allowed to dry and not quickly fixed in alcohol, drying artefact can occur, the cytoplasm will be more eosinophilic, and nuclear details will appear fuzzy.
Papanicolaou (Pap) or hematoxylin and eosin
(H&E)
58
Cytological investigation of body fluids such as urine samples, cerebrospinal fluid, pleural or peritoneal effusions, obtained by aspiration, can cast light on the ________as opposed to reactive processes.
origin and type of primary tumors
59
Serous effusion refers to the fluid accumulated in the three serous cavities namely(3), which are important sources of diagnostic material.
pleural, pericardial and peritoneal
60
The presence of malignant cells in ***serous effusions*** usually indicate_____ involvement and, therefore, a higher stage of cancer.
Collections of specimen are usually processed in the same way as a______.
metastatic
bronchial secretion
61
Jelly-like clots forming alter removal may be prevented by adding ______of Heparin for every____. of aspirate.
This is usually done by collecting the specimen in heparinized collection containers.
They can be collected in tubes or syringes that may be either plain or pre-heparinized, to prevent coagulation.
300 units
100 ml
62
Body fluids
Freshly tapped specimens are preferred for cytology.
If immediate processing is not possible, it can be preserved in the refrigerator for a period of ______ or pre-fixed in _______.
_______slides should be used to prepare smears from prefixed sample.
24-48 hours or pre-fixed in 50% ethanol.
Albuminized
63
_______preparation is by far the best method to collect cells from body fluid (e.g., urine, pleural or peritoneal fluid).
Cytospin
64
A major objection to the use of_______ is the distortion of cellular morphology due to air drying artifacts, which can be avoided by immediate fixation or by using an equal volume of polyethylene glycol.
cytocentrifuge
65
may be received as the result of direct taps of pleural or peritoneal effusions, as well as from cerebrospinal fluid (CSF) and synovial fluid.
Cell suspensions
66
Cell suspension
The optimum amount is _______ml.
Cells can remain viable for up to _____if the specimen is kept refrigerated at____ (do not freeze).
20 to 30 ml
4 days
4°C
67
Cell suspension
Urine, serous effusions and watery lavages (including bronchoalveolar lavage) all require concentration of cells prior to transferring them to glass slides._____ is the standard technique.
Centrifugation
68
Preparation of cytospin slides:
The specimen is centrifuged as soon as possible at______ for ______ (our suggested method), and the supernatant fluid is removed.
1000 RPM speed for 1 minute
69
Prep if cytospin slide
A smear is made of the sediment on a glass slide which has been previously coated with a thin layer of______, spreading the smear with another slide as evenly as possible.
When it begins to dry around the edges but is still moist in the center, the smear is fixed in 95% alcohol.
egg albumin
70
Prep of cytospin slides
Note:
When secretions are small in amount, the smear should be prepared and fixed in the operating room/bronchoscopy room to prevent specimen_____.
drying
71
Urinary Tract Specimen
Since the diagnosis is based on the type of specimens under investigation, it is extremely important to get the information about the type of specimen, and collection technique when dealing with urinary specimens.
Specimens should be clearly labeled as:
• Voided urine
• Catheterized specimen
• Washings from bladder or renal pelvis
72
Note: The_______ urine should be discarded, due to the overnight degeneration of cells.
The_____ urine is preferred.
The freshly collected urine should be sent to the lab immediately.
If delay in transportation is anticipated,_____ is recommended. We do not recommend use of any_____.
first voided
second
refrigeration
preservative
73
Voided urine from males is usually sufficient for cytological evaluation, but for female patients, a______ specimen is recommended to prevent contamination of specimen with vulvar cells.
catheterized
74
Urine
Early morning specimen yields the greatest number of cells, but such cells are usually distorted due to
prolonged bladder retention.
75
Urine
To obtain a more reliable cytological evaluation, urine specimen may have to be collected and examined____ -one in the ______and another_____.
At least_____ml is needed, which must be centrifuged.
twice
early morning; later in the day
50 ml
76
In general urine cytology alone offers a very low diagnostic yield for detection of_______.
The _____should be always mentioned on the requisition form.
The cytomorphologic features of low grade urothelial carcinomas may be indistinguishable from those of reactive urothelial hyperplasia.
urothelial carcinomas
collection technique
77
Body Cavity Effusions
The fluid specimens should be collected in a ________container, and submitted fresh to the laboratory.
clean, non-sterile dry
78
Body cavity effusion
If transportation is delayed, the specimen should be placed in the_____.
Use of_______ should be avoided.
______, on the other hand, does not interfere with cytologic preparation and evaluation.
refrigerator
Preservatives
Heparin
79
Cells in _______samples degenerate very quickly and are usually present in very low numbers.
Once the slide has been prepared, it should be rapidly air dried before staining.
cerebrospinal fluid (CSF)
80
Body cavity effusion
The addition of an equal amount of______ to the cerebrospinal fluid (CSF) is recommended if a delay in processing is anticipated.
For CSF specimens, a minimum amount of____. is necessary for cytologic evaluation.
ethyl alcohol
1 cc
81
_______fixatives are substitutes for wet fixatives.
They are either aerosols applied by spraying the cellular samples or a liquid base, which is dropped onto the slide.
Carbowax (Polyethylene Glycol)
82
The process of submerging of freshly prepared smears immediately in a liquid fixative is called_______.
This is the ideal method for fixing all _______ or ______smears
wet fixation
gynecological and non-gynecological
83
Wet fixation (5)
1. 95% Ethyl Alcohol (Ethanol)
2. Ether alcohol mixture
3. 100% Methanol
4.80% Propanol and Isopropanol
5. Denatured alcohol
84
This is the ideal fixative recommended in most laboratories for cytological specimen.
As a dehydrating agent, it causes enough cell shrinkage to yield optimal chromatin detail characteristics of cytological preparations.
95% Ethyl Alcohol (Ethanol):
85
This fixative was originally recommended by Papanicolaou.
It is an excellent fixative, but it is not used in most of the laboratories because of its safety hazards.
Ether alcohol mixture
86
Ether alcohol mixture
It consists of
equal parts of ether and 95% ethyl alcohol
87
: is an acceptable substitute for 95% ethanol.
It causes less shrinkage and is more expensive than ethanol.
100% Methanol
88
is a special purpose fixative used to hemolyze the red blood cells in hemorrhagic samples.
It is an excellent nuclear fixative as well as preservative for glycogen but results in considerable shrinkage of cells and tends to produce over staining in hematoxylin.
Carnoy's fixative
89
method is the routine staining procedure used in cytopathology laboratory.
It is a polychrome staining reaction that results in well stained nuclear chromatin, differential cytoplasmic counterstaining and cytoplasmic transparency.
Papanicolaou staining
90
Pap stain
Cytoplasmic stains are
OG-6 and EA-36.
91
synthetic stains:
OG-6 is a_______ stain while EA-36 is a______ stain.
monochrome
polychrome
92
Formalin-fixed cytology preparations must be stained with either_____ or ____
H&E or Papanicolaou stain.
93
In addition to Pap and H&E stains, many laboratories use ________ and _______ stains for cytological diagnosis of non -gynecological specimens and_____ for rapid staining.
May-Grunwald-Giemsa (MGG) and modified Wright-Giemsa
Dip Quick
94
Most_______ stains used in cytology are aqueous stains as opposed to the methyl-alcohol based stains used in hematology.
These stains are generally used for air-dried fine needle aspirates.
Romanowsky
95
The most frequently used stains for cytochemistry are:
_______ for glycogen
_____ for hemosiderin
_____ for mucins
_____ for fungal organisms
______for acid fast bacilli
Periodic Acid Schiff (PAS)
Perl's stain
Alcian Blue
Grocott Methenamine Silver (GMS)
Ziehl-Neelsen stain
96
To date, ______is considered to be the staining method of choice for exfoliative cytology.
This staining technique, known as the____, is still the gold standard today.
Papanicolaou (Pap) smear
Pap stain
97
is a multichromatic staining cytological technique that is used to differentiate cells in smear preparations of various bodily secretions, including gynecological smears, sputum, brushings, washings, urine, cerebrospinal fluid, abdominal fluid, pleural fluid, synovial fluid, seminal fluid, fine needle aspiration material, tumor touch samples, or other materials containing cells.
Papanicolaou's stain
98
EA 50 eosin yellowish
Results
Nuclei blue/black
Cytoplasm (non-keratinizing squamous cells) blue/green
Keratinizing cells pink/orange
99
Cells found in cervico-vaginal smears are usually divided into:
Mature superficial cells
Intermediate cells
Parabasal cells
100
These polygonal squamous cells measure 45-50 pm in diameter and are usually identified by the presence of pale, pink-staining cytoplasm and dark pyknotic nuclei, less than 6 u in diameter.
Mature superficial cells
101
True acidophilia is characteristic of superficial vaginal cells under_____ influence (this is not, however, a reliable index of maturation).
estrogen
102
- are medium sized polyhedral or elongated cells with basophilic vacuolated cytoplasm.
Intermediate cells
103
- are round to oval cells with small dense basophilic cytoplasm, and a total cell diameter of 15-30 pm.
Parabasal cells
104
They are smaller than intermediate cells, and have a larger vesicular nucleus.
Parabasal cells
105
Parabasal cells
They are normally found from…
two weeks of age to puberty
after childbirth
with abortions
after menopause
106
Other cells that may be found in cervico-vaginal smears include:
Navicular cells
Pregnancy cells
Endometrial cells
Endocervical glandular cells
107
are boat-shaped intermediate cells with strong tendency to fold or curl on edges.
Navicular cells
108
Their presence suggests a combined estrogen-progesterone effect.
Navicular cells
109
They are found in the latter half of the menstrual cycle, during pregnancy and menopause.
Navicular cells
110
are round, oval or boat-shaped cells with translucent basophilic cytoplasm observed greatest at the center of the cell, due to glycogen accumulation, pushing the nucleus to the side or towards the cell membrane.
This appearance is characteristic due to a deeper blue stain of the cytoplasm at the periphery.
Pregnancy cells
111
are small cells, slightly cylindrical with less basophilic cytoplasm, occurring in tightly packed groups of 3 or more.
• Endometrial cells
112
They are found during and 1-10 days after menstruation, and are shed in response to ovarian hormones
Endometrial cells
113
A smear taken during menstruation will contain_____ cells and numerous erythrocytes and leukocytes, frequently rendering the smear unsatisfactory for a reliable assessment.
endometrial cells
114
occur in large groups or small sheets.
The cytoplasm is usually stained pale blue/gray and is finely vacuolated, often with indistinct cell borders and nuclei with finely granular chromatin.
• Endocervical glandular cells
115
They may present a honeycomb appearance when viewed on end.
Endocervical glandular cells
116
, aside from being relatively inexpensive, may be performed regularly without undue risk, even in pregnant women.
Vaginal smears prepared for such purpose are best taken from the _____of the vaginal wall, because it is more accessible and less likely to be contaminated by cellular debris or vaginal discharges.
Vaginal hormonal cytology
upper lateral third
117
Staining Procedure for Non-Gynecologic Specimens:
We use a_______ to provide a standardized method for staining non-gynecologic specimens.
_______is the optimum nuclear stain and the combination of OG-6 and EA 50 gives the subtle range of green, blue and pink hues to the cell cytoplasm.
modified Papanicolaou technique
Harris' hematoxylin
118
This procedure provides optimum nuclear detail information, which is important for cytologic evaluation of Pap smears, nongynecological and FNA cytologic specimens.
Modified Papanicolaou Staining
119
This is one of the common Romanowsky stains used in cytology. It is useful for studying cell morphology in air-dried smears.
May-Grunwald Giemsa Stain:
120
It is superior to Papanicolaou stain when studying the cytoplasm, granules, vacuoles, and basement membrane material.
May-Grunwald Giemsa Stain
121
For nuclear staining_____ stain is superior.
Papanicolaou stain
122
Mounting
Long-term specimens are mounted with mounting media that are first applied to the slide in dissolved form and then harden as the solvent evaporates.
The refractive index of mounting media is approximately_____ and is close to that of glass.
For optimum optical properties, transparency and brilliance of specimens it is important to ensure that the mounting medium contains the solvent that was used for clearing.
1.5
123
The advantages of mounting agents containing xylene or toluene include:
• Optical brilliance
• Good drying times
• No air bubbles under the coverslip
• Color stability of stained slides
124
Immunohistochemical studies for_______ on cell smears, imprints, and cytocentrifuge preparations show sensitivity equal to, or greater than, that of histologic sections.
By contrast,_________ may be more difficult to demonstrate in cytologic material, probably because cells remain structurally intact and do not allow for easy penetration of immunologic reagents.
cell surface antigens
intracytoplasmic and intranuclear antigens
125
IHC
Samples stored for several months, either at room temperature,____ or
____can be processed for DNA and protein analysis.
4°C; -20°C
126
IHC
_______and subsequent direct sequencing can be used to analyze mutations of p53 and other genes from relatively few suspicious or malignant cells.
Polymerase chain reaction (PCR)
127
IHC
is the most objective technique for assessment of Her2/neu of fine needle aspirates for assessment of breast cancer.
Fluorescence In Situ Hybridization (FISH)
