HISTOPATHOLOGY FINALS Flashcards
Deals with microscopic study of cells desquamated from epithelial cells
Exfoliative cytology
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
Specimens for Examination: Non-gynecological
Sputum
Smears of urine sediments
Prostatic and breast secretions
Pleural and peritoneal fluids
Gastric and bronchial secretions
CSF
Specimens for Examination: Gynecological
Endometrial and Endocervical smears
Cervico vaginal smear a.k.a PAP Smear
Smear to detect HPV, Trichomoniasis, Yeast infection
PAP smear (Cervicovaginal smear)
Preparation for NON-GYNECOLOGIC SPECIMENS
Smears
Cell blocks
Cytospin preparation
Motion for streaking smear
Zigzag
Smear applied at the center and use loop to spread
Spreading
Smear technique for thick secretion
Pull apart technique
Smear technique for fixation
Touch preparation/ Impression smear
Best method to collect cells from body fluids
Cell blocks
Cytospin preparation rpm and minutes
1000 rpm for 1 minute
Specimen to rule out pneumocystis jirovecci
BAL (Bronchoalveolar lavage)
Respiratory tract specimens
Bronchial washings
Bronchial brushings
Sputum
Brochoalveolar lavage
Manner of collection for sputum
Collected from a deep cough
At least 3 mornings (3 days)
Cells that indicate that the specimen is sputum
Alveolar macrophage
Fixative for sputum
Saccomano fluid
Fixative for bronchial brushing
spray fixative or 95% ethanol
Gastric secretions/aspirates should not be delayed beyond ____ minutes to prevent degradation
30 minutes
Requirement for gastric secretion/aspirate collection
Fasting for at least 8 hours
Smear preparation for breast secretion
pull apart technique
Discharge from nipple is usually benign and may be due to lesion like
duct ecstasia
papilloma
Peritoneal-Pleural-Pericardial fluid presence is always —
pathologic
Jelly clots in peritoneal-pleural-pericardial fluid is prevented by adding —-
300 units of heparin per 100mL of aspirate
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
Concentrated Sputum
Urinary sediments
Bronchial labage
Specimens with trypsin
Adhesives for cytology
Celloidin ether alcohol
Leuconostoc culture - bacteria, gram positive cocci
APES – 3 aminopropyltriethoxysilane
Pooled 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