CYTOLOGIC TECHNIQUES Flashcards by Kyle Ancheta

_____________ and ____________ are most important in cytological interpretation

consistency and reliability
- cytologists rely heavily on the quality and appearance of the stain

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Deals with the microscopic study of cells that have been desquamated from epithelial surfaces

Exfoliative Cytology
- exfoliated cells may be found in smears that have been spontaneously shed or physically removed from epithelial and mucous membranes

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Commonly received cytoprep

Thin Prep
➢ wet-fixed in 95% ethanol immediately after preparation to preserve the fine structure of the chromatin and help in the evaluation of nuclear changes
➢ air drying is avoided

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The commonest cancer that can be detected even at the pre-invasive stage

cancer of the uterine cervix

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A cervical cytology screening involves the microscopic examination of cell samples that have been taken primarily from the ecto- and endocervix

Pap Smear

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True or False
- cervical cytology is a diagnostic test and abnormal findings must be confirmed histologically

False
- screening test

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True or False
- patient should abstain from coitus, not douche the vagina for at least a day, and not apply intravaginal preparations for at least one day before the examination

False
- not apply intravaginal preparations for at least one week before the examination

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True or False
- smears should not be taken during menstrual bleeding

True
- contamination with blood, endometrial component, and tissue debris that can obscure the cells during smear examination

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True or False
- Cotton swabs are discouraged because of the drying artifacts and loss of cells

True

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Preferable to plastic spatula because of its mildly rough
surface that can collect more material

Wooden spatula

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True or False
- excessively thin or thick smears can result in false-negative reports

True

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An exfoliative cytology technique used for ulcerated surface lesions to allow an immediate assessment of the lesion before fixation and processing of the tissue sample

Impression smear
- preparations should be quickly air dried and then stained
- indicated in the case of tumors especially of lymph nodes

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For sputum smear, obtain at least _ consecutive morning sputum specimens

at least 3 consecutive morning sputum specimens

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Agent use in early morning sputum by a deep cough

Saccomanno fluid

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Composition of Saccomanno fluid

50% ethyl alcohol and 2% carbowax

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True or False
- for more extensive study in sputum, it is recommended that a minimum of 2-4 slides be prepared, and one is air dried for Giemsa staining; at least two slides should be stained by Papanicolaou method

True

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Absence of this cell suggests “saliva” rather than sputum and results an unsatisfactory specimen

alveolar macrophages

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True or False
- For smears of gastric secretions
and aspirates, any delay of more than 1/2 hour, before fixation, will digest the cells and make the specimen unsatisfactory for evaluation

True

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For smears of gastric secretions
and aspirates, patient should have fasted for at least _ hours before gastric washing is performed

at least 8 hours

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True or False
- In women, any discharge from the nipple is abnormal

True
- except during lactation and the
immediate post-lactation period

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True or False
- cytologic examination of nipple discharge has an extremely high diagnostic yield for diagnosis of breast carcinoma

False
- extremely low diagnostic yield

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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 (FNA)

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Cytologic examination that 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 (FNA)

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FNA basic technique uses a __-gauge needle and a __-ml syringe

25-gauge needle and a 10-ml syringe

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True or False - FNA sample should be air dried as quickly as possible to reduce the effects of shrinkage

True

True or False - For FNA slide preparation, when a solid lesion is aspirated, usually a few drops from the tip of the needle has the least diagnostic material for cytologic evaluation

False - most diagnostic material

Recommended slide to be prepared from FNA sample

prepare a maximum 4 slides

FNA ideal aspirate is ________ consistency with numerous cells suspended in a small amount of tissue fluid without a mixture with blood

creamy consistency

Smears obtained by FNA for hematological stains (such as May–Grunwald–Giemsa) are fixed through?

air dried

Smears obtained by FNA for Papanicolaou (Pap) or hematoxylin and eosin (H&E) should be fixed through?

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

Jelly-like clots obtain from body fluids forming alter removal and prevented by adding _____________

300 units of Heparin for every 100 ml of aspirate - usually done by collecting the specimen in heparinized collection containers

Describe a preferred specimen for cytology

Freshly Tapped Specimens

Albuminized slides should be used to prepare smears from __________ sample

prefixed sample

By far the best method to collect cells from body fluid (e.g., urine, pleural or peritoneal fluid)

Cytospin preparation

True or False - Suggested method for cytospin preparation is where specimen is centrifuged as soon as possible at 1000 RPM speed for 5 minute

False - 1000 RPM speed for 1 minute

Received as the result of direct taps of pleural or peritoneal effusions, as well as from cerebrospinal fluid (CSF) and synovial fluid

Cell Suspensions - centrifugation is the standard technique

Cell suspension optimum amount is __ to __ ml and cells can remain viable for up to _ days if the specimen is kept refrigerated at _°C (do not freeze)

Cell suspension optimum amount is 20 to 30 ml and cells can remain viable for up to 4 days if the specimen is kept refrigerated at 4°C (do not freeze)

True or False - For cytologic examination of urinary tract specimen, first voided urine is preferred and the second urine is discarded

False - first voided urine should be discarded due to the overnight degeneration of cells, second urine is preferred

True or False - Using preservative for urine specimen for cytology is not recommended if delay in transportation

True - If delay in transportation is anticipated, refrigeration is recommended. Using preservative is not recommended.

True or False - Catheterized specimen is recommended for male and female in urine cytology study

False - voided urine from males is usually sufficient for cytological evaluation, but for female patients, a catheterized specimen is recommended to prevent contamination of specimen with vulvar cells

True or False - to obtain a more reliable cytological evaluation, urine specimen may have to be collected and examined twice (one in the early morning and another later in the day)

True - at least 50 ml is needed, which must be centrifuged

True or False - Heparin does not interfere with cytologic preparation and evaluation

True

Distance when sprayed fixative is use on a smear

about a foot distance

____________ prevents the slide from sticking together in the fixative

paper clip

True or False - if smears cannot be made immediately, the collected material should be placed in 50% alcohol for all types of effusions

True - can be replaced by Saccomano preservative (50% alcohol and carbowax)

Ideal fixative recommended in most laboratories for cytological specimen

95% Ethyl Alcohol (Ethanol) - as a dehydrating agent, it causes enough cell shrinkage to yield optimal chromatin detail characteristics of cytological preparations

Originally recommended by Papanicolaou and it consists of equal parts of ether and 95% ethyl alcohol, an excellent fixative but not used in most of the laboratories because of its safety hazards

Ether alcohol mixture

An acceptable substitute for 95% ethanol. It causes less shrinkage and is more expensive than ethanol

100% Methanol

Process of submerging of freshly prepared smears immediately in a liquid fixative and ideal method for fixing all gynecological and non-gynecological smears

Wet fixation

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) fixatives

A special purpose fixative used to hemolyze the red blood cells in hemorrhagic samples

Carnoy’s fixative

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 - must be prepared fresh when needed and discarded after each use

Routine staining procedure used in cytopathology laboratory

Papanicolaou staining method

A polychrome staining reaction that results in well stained nuclear chromatin, differential cytoplasmic counterstaining and cytoplasmic transparency

Papanicolaou staining method - method of choice for exfoliative cytology - gold standard

True or False - Nuclear stains are OG-6 and EA-36; both are synthetic stains

False - Cytoplasmic stains

True or False - OG-6 is a polychrome stain while EA-36 is a monochrome stain

False - OG-6 is a monochrome stain while EA-36 is a polychrome stain

Use for cytological diagnosis of non-gynecological specimens

May-Grunwald Giemsa (MGG) and modified Wright-Giemsa stains

Use for rapid staining

Dip Quick

Stain used in cytology that are mostly aqueous stains as opposed to the methyl-alcohol based stains used in hematology; used for air-dried fine needle aspirates

Romanowsky stains

Most Frequently Used Stains for Cytochemistry: a) Periodic Acid Schiff (PAS) for __________ b) Perl’s stain for _______________ c) Alcian Blue for ___________ d) Grocott Methenamine Silver (GMS) for ___________________ e) Ziehl-Neelsen stain for ________________

Most Frequently Used Stains for Cytochemistry: a) Periodic Acid Schiff (PAS) for glycogen b) Perl’s stain for hemosiderin c) Alcian Blue for mucins d) Grocott Methenamine Silver (GMS) for fungal organisms e) Ziehl-Neelsen stain for acid fast bacilli.

Enzymes may be used for immunohistochemistry

peroxidase and alkaline phosphatase

A multichromatic staining cytological technique that is used to differentiate cells in smear preparations of various bodily secretions

Papanicolaou's stain (also known as Pap stain)

Five dyes used in Pap's stain

1. Hematoxylin 2. OG-6 3. EA 4. Light Green SF yellowish 5. Bismarck brown Y

Dye in Pap's stain that is used to stain the nuclei

Hematoxylin

First counterstain used in Pao's stain

OG-6

-6 in OG-6 counterstain denotes what?

-6 denotes the used concentration of phosphotungstic acid

___________ stains keratin and was originally used to stain the small cells of keratinizing squamous cell carcinoma present in sputum

Orange G

Second counterstain used in Pap's stain

EA (Eosin Azure) - comprising three dyes; the number denotes the proportion of the dyes, e.g. EA-36, EA-50, EA-65

_________ stains the superficial epithelial squamous cells, nucleoli, cilia, and red blood cells

Eosin Y

__________________________ stains the cytoplasm of other cells, including non-keratinized squamous cells

Light Green SF yellowish

Can be used as an alternative stain to light green SF yellowish, but it produces visually different results

Fast Green FCF

Stains nothing and often omitted

Bismarck brown Y

Stained specimen should display hues from the entire spectrum: ➢ _________ are crisp blue to black ➢ cells with high content of keratin and glycogen are _________ ➢ _________________ are orange to pink ➢ intermediate and parabasal cells are __________________ to _____ ➢ metaplastic cells often stain both _______ and ______ at once

Stained specimen should display hues from the entire spectrum: ➢ cell nuclei are crisp blue to black ➢ cells with high content of keratin and glycogen are yellow ➢ superficial cells are orange to pink ➢ intermediate and parabasal cells are turquoise green to blue ➢ metaplastic cells often stain both green and pink at once

True or False - Hematoxylin stains lose strength more rapidly than OG and EA, and should be replaced every week or as soon as the staining of cells are no longer sharp and crisp

False - OG and EA stains lose strength more rapidly than hematoxylin and should be replaced every week or as soon as the staining of cells are no longer sharp and crisp

Original Pap's Staining Method in order

1. 96% ethyl alcohol 2. 70% ethyl alcohol 3. 50% ethyl alcohol 4. Distilled water 5. Harris hematoxylin 6. Distilled water 7. 0.5% HCl acid 8. Distilled water 9. 0.1% ammonia water 10. 50% ethyl alcohol 11. 70% ethyl alcohol 12. 96% ethyl alcohol 13. OG-6 14. 96% ethyl alcohol 15. 96% ethyl alcohol 16. EA-50 eosin yellowish 17. 96% ethyl alcohol 18. 100% ethyl alcohol 19. Xylene 20. Mount: in DPX using coverslip

Polygonal squamous cells that measure 45-50 µm in diameter and are usually identified by the presence of pale, pink-staining cytoplasm and dark pyknotic nuclei, less than 6 µ in diameter

Mature Superficial Cells

Characterize by true acidophilia under estrogen influence (not a reliable index of maturation)

Mature Superficial Cells

_____________________ may be observed due to the drying of smears especially before fixation, prolapse and drying of vaginal epithelium, infection and chemicals

Pseudo-acidophilia

Medium sized polyhedral or elongated cells with basophilic vacuolated cytoplasm

Intermediate Cells

Round to oval cells with small dense basophilic cytoplasm, and a total cell diameter of 15-30 µm

Parabasal cells - smaller than intermediate cells, and have a larger vesicular nucleus

Normally found from two weeks of age to puberty, after childbirth, with abortions and after menopause

Parabasal cells

Boat-shaped intermediate cells with strong tendency to fold or curl on edges

Navicular cells

Presence suggests a combined estrogen-progesterone effect

Navicular cells

Found in the latter half of the menstrual cycle, during pregnancy and menopause

Navicular cells

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

Pregnancy cells - appearance is due to a deeper blue stain of the cytoplasm at the periphery

Small cells, slightly cylindrical with less basophilic cytoplasm, occurring in tightly packed groups of 3 or more

Endometrial Cells

Found during and 1-10 days after menstruation, and are shed in response to ovarian hormones

Endometrial Cells

Occur in large groups or small sheets and 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

Present a honeycomb appearance when viewed on end

Endocervical Glandular Cells

Vaginal discharge is best taken from the ___________________ of the vaginal wall

upper lateral third of the vaginal wall - more accessible and less likely to be contaminated by cellular debris or vaginal discharges

Quantitative evaluation of vaginal smear is taken under what objective?

under 40x objective

Non-Gynecologic Specimens is performed on __________________________________ to provide a standardized method for staining nongynecologic specimens

modified Papanicolaou technique

True or False - smears should not be allowed to dry in between any of the staining steps

True

Superior to Papanicolaou stain when studying the cytoplasm, granules, vacuoles, and basement membrane material

May-Grunwald Giemsa Stain ➢ Papanicolaou stain is superior in nuclear staining

Refractive index for mounting media for cytology

approximately 1.5 (close to glass)

True or False - it is important to ensure that the mounting medium contains the solvent that was used for clearing for optimum optical properties, transparency and brilliance of specimens

True

True or False - false negative immunoperoxidase results for intracellular antigens are more common in histologic samples than in cytologic specimens

False - false negative immunoperoxidase results for intracellular antigens are more common in cytologic samples than in histologic specimens

CYTOLOGIC TECHNIQUES Flashcards

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