Papanicolau Staining Flashcards
CYTOLOGY
highly specific and accurate technique used to differentiate_____ from other_____ or _____ disease
tumors
inflammatory or degenerative
suitable for screening purposes
Cytology
Advantages of cytology
ease of obtaining material
ease of processing
Pap Smear
developed by _______ in _____
Georgios Nikolaou Papanikolaou (George Papanicolaou)
1883-1962
Georgios Nikolaou Papanikolaou (George Papanicolaou)
Father of..
Modern cytology
a test in gynecological cytology
Pap smear
used for detection and prevention of cervical cancer and other cytologic disease of the female reproductive system
Pap smear
True or False:
Pap smear involves monochromatic staining of cytologic smear
False
multichromatic
Pap smear
demonstrates ____ and _____in various stages of maturation
same technique may be used to check____ in other specimen types
vaginal and cervical cells
malignancy
True or False
Pap smear is a reliable technique
True
used to differentiate cells in smear preparations of various bodily secretions
PAPANICOLAOU STAIN
Bodily secretions used in papanicolau staining
gynecological smears
sputum
brushings and washings
urine
CSF
seminal fluid
serous fluids
synovial fluid
FNA
tumor touch samples
Composition of pap stain
Hematoxylin - primary stain
OG-6 (Orange G) Counterstain
EA (Eosin Azure) counterstain
What is the primary stain in pap stain
Hematoxylin
What are the counterstains in pap stain
OG-6 (Orange G) Counterstain
EA (Eosin Azure) counterstain
Composition of EA
Eosin Y
Light Green SF yellowish
Bismarck Brown Y
nuclear stain; it is used to stain cell nuclei (Blue or Black)
Hematoxylin
What gives the color yellow to glycogen
Hematoxylin
responsible for the yellow color imparted to glycogen
‘unmordanted’ hematein
-6 in OG-6 denotes the used concentration of
phosphotungstic acid
OG-6 stains…
Keratin
original role was to stain small cells of keratinizing squamous cell carcinoma (sputum)
OG-6
This composition of pap stain contains 3 dyes
Eosin Azure counterstain
EA counterstain
contains two mutually incompatible chemicals:
Bismarck brown
PTA (phosphotungstic acid)
precipitates each other, impairing the useful life of the mixture and compromising the differential staining of eosin and light green.
Bismarck brown and PTA (phosphotungstic acid)
stains the superficial epithelial squamous cells, nucleoli, cilia, and red blood cells.
Eosin Y
yellowish stains the cytoplasm of other cells, including non-keratinized squamous cells
Light Green SF
This dye is now quite expensive and difficult to obtain, therefore some manufacturers are switching to Fast Green FCF, however it produces visually different results and is not considered satisfactory by some.
Light Green SF
stains nothing and in contemporary formulations it is often omitted
Bismarck brown Y
True of False
Pap stain stained specimen displays hues from entire spectrum
True
Red-violet
True of False
Pap stain…
chromatin patterns well-visible
cells from borderline lesions are easier to interpret
True
True or False
Pap stain…
better photomicrographs
cells are very transparent
True
Cell nuclei -
BLUE TO BLACK
Keratin and Glycogen ->
YELLOW
Superficial cells ->
ORANGE TO PINK
Intermediate to Parabasal cells ->
TURQUOISE GREEN TO BLUE
Metaplastic cells ->
often both GREEN AND PINK
Reagen(WHO)
Class 1
Negative
Reagen(WHO)
Class 2
Inflammation
Reagen(WHO)
Class 3
Mild dysplasia
Reagen(WHO)
Class 4
Moderate dysplasia
Severe dysplasia
Carcinoma in situ
Reagen(WHO)
Class 5
Invasive cancer
Ruchart
Class 1
Negative
Ruchart
Class 5
Invasive cancer
Bethesda
Class 1
Within normal
Bethesda
Class 2
Atypical squamous cells of undetermined significance ( ASCUS)
Bethesda
Class 3
low-grade squamous intraepithelial lesion
Bethesda
Class 4
High-grade squamous intraepithelial lesion.
Bethesda
Class 5
Invasive cancer
PAPANICOLAOU STAIN
The problem…
not standardized; formulation varies among different manufacturers (do not mix from different brands)
several modifications, subtly differs in dyes, ratios and timing of process
The Cytocolor® Method is by what brand?
Merck Millipore International
The Cytocolor® Method
(Merck Millipore International)
staining time:
3 minutes
These stains provide complete information on dignity and additional information on hormone status and vaginal flora in gynaecological smears
Cytocolor method
The difference between these and classical Papanicolaou stains is that Cytocolor®…
requires no orange stain
In cytocolor method
both mature and keratinized cells appear as____ instead of orange
pink
In traditional papanicolau staining method…
Fix smears in _____ for ____ mins for fixation.
95% ethanol for 10 minutes
For cytocolor method
Fix smears with ______ or _____
Merckofix or in 95% ethanol (10 dips)
What type of hematoxylin is used in traditional pap staining
Harris hematoxylin (6mins)
Commonly used fixative in cell cytology
95% ethanol
Best fixative for cell cytology
95% ethanol ether
Flmmable
95, 75, 50% ethyl alcohol
Rehydration
Purpose of HCl 0.5% solution
Differentiator/ Decolorizer
Bluing agent in traditional method
Ammonium hydroxide
50, 70, 95% ethyl alcohol
Dehydration
Last xylene
Clearing
High refractive index
Stain in cytocolor method
Polychromic solution
Alcohol used in cytocolor
2-propanol