HISTOPATHOLOGY FINALS Flashcards by PATRICIA ONG

Deals with microscopic study of cells desquamated from epithelial cells

Exfoliative cytology

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Purpose of exfoliative cytology

Detect cancerous /malignant conditions

Detect asymptomatic cancer in women or pre-cancerous cervical lesions

Determination of genetic sex

Detection of possible infection

Evaluate female hormonal status in case of sterility

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Specimens for Examination: Non-gynecological

Sputum

Smears of urine sediments

Prostatic and breast secretions

Pleural and peritoneal fluids

Gastric and bronchial secretions

CSF

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Specimens for Examination: Gynecological

Endometrial and Endocervical smears

Cervico vaginal smear a.k.a PAP Smear

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Smear to detect HPV, Trichomoniasis, Yeast infection

PAP smear (Cervicovaginal smear)

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Preparation for NON-GYNECOLOGIC SPECIMENS

Smears
Cell blocks
Cytospin preparation

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Motion for streaking smear

Zigzag

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Smear applied at the center and use loop to spread

Spreading

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Smear technique for thick secretion

Pull apart technique

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Smear technique for fixation

Touch preparation/ Impression smear

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Best method to collect cells from body fluids

Cell blocks

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Cytospin preparation rpm and minutes

1000 rpm for 1 minute

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Specimen to rule out pneumocystis jirovecci

BAL (Bronchoalveolar lavage)

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Respiratory tract specimens

Bronchial washings

Bronchial brushings

Sputum

Brochoalveolar lavage

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Manner of collection for sputum

Collected from a deep cough

At least 3 mornings (3 days)

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Cells that indicate that the specimen is sputum

Alveolar macrophage

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Fixative for sputum

Saccomano fluid

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Fixative for bronchial brushing

spray fixative or 95% ethanol

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Gastric secretions/aspirates should not be delayed beyond ____ minutes to prevent degradation

30 minutes

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Requirement for gastric secretion/aspirate collection

Fasting for at least 8 hours

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Smear preparation for breast secretion

pull apart technique

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Discharge from nipple is usually benign and may be due to lesion like

duct ecstasia
papilloma

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Peritoneal-Pleural-Pericardial fluid presence is always —

pathologic

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Jelly clots in peritoneal-pleural-pericardial fluid is prevented by adding —-

300 units of heparin per 100mL of aspirate

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Breast secretion fixative:

spray fix or 95% Isopropyl alcohol

Used to detect urothelial malignancies

Urinary tract sediments

Urinary tract specimens

Voided urine Catheterized urine Washing from bladder or renal pelvis

Preferred urine collection for urinary tract specimen

secondary collection

Urine specimen should be ____ if there is delay in examination

refrigerated

Urine specimen volume for best results

50 mL

Body cavity effusions- CSF and ascitic fluid should be submitted _____

fresh

Amount of CSF should be ----

at least 1 mL

Extra sediments are used for -----

cell block technique

Methods of Cell block preparation

Direct filtration Plasma thrombin method Carbowax method

Method of Cell block preparation: Centri fluid, add 10% formalin to sediment, embed using paraffin

Direct filtration

Method of Cell block preparation: (not usually carried out, expensive) - Centri fluid, add 1 ml of plasma and add thrombin

Plasma thrombin method

Disadvantage of direct filtration

easily washed out

Advantage of plasma thrombin method

reduce washing out

Advantage of carbowax method

no dehydration step but is expensive

Smears should be prepared from fresh material and must be prepared and FIX while moist within

10 - 15 minutes

Specimens that require adhesives

**C**oncentrated Sputum **U**rinary sediments **B**ronchial labage **S**pecimens with trypsin

**Adhesives for cytology**

**C**elloidin ether alcohol **L**euconostoc culture - bacteria, gram positive cocci **A**PES – 3 aminopropyltriethoxysilane **P**ooled serum or plasma

Most common tissue adhesive

Mayer's egg albumin

Egg albumin is not used in cell cytology due to ---

intensely stained by counterstain in PAP's

Best Fixative for Cytology

95% ethanol with ether (not commonly used, flammable)

Commonly used fixative for cytology

95 % ethanol

Spray fixative distance from the slide

1 foot (12 inches or 30.5 cm)

Saccomano Preservative components

Combination of 50% alcohol and 20% carbowax

Fixation of fluid specimens: percentage of alcohol is for all types of effusions

50%

Fixation of fluid specimens: peritoneal and pleural fluid

50%

Fixation of fluid specimens: percentage for sputum specimen

70%

Fixation of fluid specimens: percentage for urine, gastric, and bronchial aspirates

95%

The gold standard stain for cytology

Papanicolau stain

Components of PAP's stain

Harris Hematoxylin OG6 EA 50

Component of PAP's stain for nucleus

Harris hematoxylin

PAP's stain: Counterstain for cytoplasm of superficial cells

OG6

PAP's stain: Counterstain for both parabasal and intermediate cells

EA 50

Best for bloody stains; cytoplasm green

Carnoy's fluid

Color result: Vesicular Nucleus

Blue

Color result: Pyknotic nucleus

dark blue to black cytoplasm

Color result: OG 6

orange with a hint of green

Color result: EA36-50

olive green with a hint of brown and red

Color result: Bacteria

Dark blue

Color result: Mycelia

Violet

Color result: T. vaginalis

pale greenish blue

Purpose of PAP smear

Screening test for cervical cancer For hormonal evaluation - steroid hormones, vaginal cells

PAP's smear method: causes crowded cells, specimen is directly placed on the slide

Conventional PAP's

Used to collect PAP smear

ayres spatula

Pap smear method: Use of spatula or brush/broom to collect specimen. Placed in a vial containing preservative. A thin layer of cells is placed on a slide

Liquid based PAP's

Sites of collection PAP: For evaluating inflammatory conditions

Upper third of the vaginal wall

Sites of collection PAP: Detection of lesions or intrauterine lesions

Endocervix

Sites of collection PAP: most common site for cancer screening

Ectocervix

Ectocervix is lined up with

stratified squamous non keratinized

Endocervix is lined up with

simple columnar epithelium

Location for PAP smear preparation; junction between endo and ectocervix

Transformation zone (T zone)

Hormone that influence desquamation of superficial cells

Estrogen

Estrogen most predominant in menopausal

E1 - Estrone

Estrogen predominant in reproductive years

E2 - Estradiol

Estrogen most plentiful, least potent

E3 - Estriol

Hormone that influence desquamation of intermediate cells

Progesterone

Smears for hormonal cytology should be taken from the | for PAP smear

upper lateral third of vaginal wall

Cells in PAP's: Largest and most mature with pyknotic nuclei (smallest) 30-60u Polyhedral flat cells and cytoplasm may be acidophilic or basophilic Pink cytoplasm

Superficial cells

Cells in PAP's: Medium sized cells 20-30 u Polyhedral cells with basophilic cytoplasm with vacuoles

Intermediate cells

Intermediate cell, boat shaped cells with a tendency to fold or curl on edges.

Navicular cells

Cells in PAP's: Found in the latter half of menstrual cycle, during pregnancy or menopause May suggest progesterone-estrogen effect May be found as a result of abnormal androgen stimulation

Intermediate cells

Cells in PAP's: Round to oval (15-30 u) With strongly basophilic cytoplasm Found from 2 wks of age to puberty, after childbirth, abortion and after menopause

Parabasal cells

Cells in PAP's: Smallest and not normally shed 13-20 u Round to slightly oval cells with large nucleus occupying half or more of the cell volume, with strongly basophilic cytoplasm Found only before pregnancy and ff> menopause

Basal cells

Cells in PAP's: Found during and 1-4 days after menstruation Occurring in groups of 3 or more shed in response to ovarian hormone If seen in post menopausal women it may indicate possible endometrial carcinoma or endometrial hyperplasia

Endometrial cells

Cells normally shed in vagina:

Superficial cells Intermediate cells Basal cells

Occurring in large groups or sheets Forms the characteristic honey comb appearance

ENDOCERVICAL GLANDULAR CELL CARCINOMA

Abnormal Cellular Components in PAP’s: seen in diabetic patients, in those taking oral contraceptives, patients under prolonged steroid therapy and among immunocompromised patients

Candida albicans (yeast)

Abnormal Cellular Components in PAP’s: Pear shaped parasite

T. Vaginalis (trophozoite)

Abnormal Cellular Components in PAP's: Indicative of bacterial vaginosis

Clue cells (squamous cells with attached bacilli)

Abnormal Cellular Components in PAP's: Abnormal squamous with atypical nucleus surrounded by non-staining halo, indicative of HPV infection

Koilocytes

Abnormal Cellular Components in PAP's: Doderlain bacillus is increasedly seen in

Last trimester of prengancy Infection Estrogen deficiency Diabetes Mellitus

Involves evaluation of hormonal status based on the distribution of cells

Cytohormonal Maturation Index (CHMI)

CHMI: Parabasal, Intermediate and Superficial cells result 0 / 90 / 10

Newborns

CHMI: Parabasal, Intermediate and Superficial cells result: 0 /80 / 20

Menopause

CHMI: Parabasal, Intermediate and Superficial cells result: 0 / 70 / 30

Post ovulatory

Old method, obsolete manner of reporting PAP’s

Class system

Class system result: Class I

Negative malignancy

Class system result: Class V

Conclusive of malignancy

New system of reporting cervical, vaginal cytologic smears

Bethesda system

Bethesda system is developed at

National Cancer Institute December 1988

Report format for specimen adequacy:

satisfactory limited unsatisfactory

Detect cause of death

Autopsy

method to detect cancerous condition

Biopsy