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Significance of Exfoliate Cytology

HISTOCYTOLOGIC 1. For assessing cancerous conditions
2. For assessing female hormonal
TECHNIQUE activity
3. For determination of genetic sex
Prepared by: EGB, RMT 4. To differentiate cancer from other
diseases like infection, inflammation
Histopathologic technique and others
• Involves the different procedures that have 5. To determine the different cytologic
been adopted for the preparation of indices
materials and tissues for microscopic
examination whether these tissues are Hormonal Index/Hormonal Statute (HI)
normal (histologic) or abnormal
• Ratio between gonadotropic hormone of
(histopathologic)
a person
Scope of Study • In male- testosterone/androgen
• In female- estrogen/progesterone
1. Cytologic Significance of Hormonal Index
2. Histopathologic 1. To determine if the reproductive system
is functioning
Cytologic technique
2. To determine the fertility of sterility
• Preparation of cells in form of smears for
3. To diagnose tumor especially cancer
microscopic diagnosis
Laboratory diagnosis of HI
• In relation with cytology (study of normal 1. Smear preparation
cells) and cytopathology (study of 2. Direct microscopic examination
abnormal cells) Result of laboratory diagnosis of HI
Exfoliative Cytology 1. Formation of pale-leaf or forming crystal
• Study of cells that are removed off from is a sure sign of fertility
different parts of the body which o Estrogen Progesterone- fertility
includes the following: o Progesterone Estrogen- sterility
1. Lining epithelium Cancer in HI
2. Mucosal surfaces • There is persistent high level of hormone
3. Fluid aspirates (estrogen/progesterone) when examined
for 28 consecutive days; same case as in
• Examples: male
Female scrapings Vagina
Cervix
Uterus Cytologic Indices
Male Prostatic tissue Maturation Index (MI)
Respiratory organ Sputum and bronchial • Comparison between the number of
washing immature and mature cells and
GIT Gastric and duodenal determine the location and extent of the
(Gastrointestinal fluid disease in the body
tract) Significance of MI
Bone marrow 1. Increase in cases of cancer
CSF 2. Increase in cases of abnormal hormone
(Cerebrospinal Pyknotic Index (PI)
fluid) • Percentage of cells with small, shrunken,
Body cavity Transudate structureless nuclei
Exudate Significance of PI
Skin scrapings 1. Increase in cases of cancer
2. Increase in cases of necrosis on
degeneration

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Basophilic Index/Acidophilic Index (AI) Millipore Filter Technique

• Percentage of cell staining capacity • same method or procedure as in Pap’s
Significance of AI smear but differs in sedimentation
1. Normal staining of cytoplasm is acidic instead it uses a special filter apparatus
Vaginal Index (VI) for ultrafiltration for collecting
• Comparison of the different types of sediments
cells Pre-cone or Mini-cone
Types of Cells are: • A special procedure which is done by a
1. Mature superficial cells direct biopsy with pre-cone apparatus to
2. Intermediate cells collect the specimen, that is well fitted
3. Parabasal cells to the female cervix
4. Basal cells • Its purpose is to screen the possibility of
Significance of VI cancer of the cervix
1. To determine the location and extent of • If in this method no cancer cells are seen,
disease an alternative method is Pap’s smear
2. To differentiate cancer from infection • If it is still negative to Pap’s smear the
3. To detect pregnancy last resort is Surgical biopsy which is the
punch biopsy
Punch biopsy
Important Laboratory Methods in Cytology
• A special biopsy method for cervix
Papanicolau/Pap’s smear
• Said to be the best method of collection
• Routine cytologic procedure
Collection of specimens
• Most common method used in cytology Female specimen By simple vaginal
Pap’s staining procedure scrapings aspiration
1. Wash the fixed smear in water
Male specimen Prostatic massage
2. Dip in descending ethyl alcohol
Bronchial washing By aspiration of fluid
o Done for rehydration
known as
o Ex: 95%, 70%, 50% EA
Bronchoscopy and
3. Wash again with water
the apparatus used is
4. Dip in Harris Hematoxylin for 3-5 mins.
Bronchoscope
o Serve as the primary or basic
Gastric/Duodenal By nasogastric
stain
fluid intubation
5. Wash with water thoroughly (usually 3
changes)
Purpose is to detect
6. Dip in Acid alcohol and wash with water
GIT cancer
immediately
o Serve as the decolorizer
If the test for reaction
7. Dip in Ammonia Water three time (3x)
is:
and wash with water immediately
-Acidic- from gastric
8. Dip in ascending ethyl alcohol
-Basic- from duodenal
o For dehydration
Bone marrow Puncture is done
9. Dip in OG6 for 2 mins
commonly in ilium by
o Serve as intensifier
using a needle
10. Wash by means of ethyl alcohol
CSF (Cerebrospinal By lumbar tap
11. Dip in EA50 for 3-5 mins.
fluid)
o Serve as the acidic counterstain
12. Wash with ethyl alcohol thoroughly Transudates and Needle puncture
13. Dip in Xylol for few mins. Exudates aspiration can from:
o Serve as clearing agent - Lungs- pleural fluid
Cell Block Method - Heart- pericardial
fluid
• A combination of cytotechnique and
- Abdomen- ascitic
histopathologic technique
fluid

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skin By simple scrapings Fuelgen reaction

Wet Film Method Utilizing supravital
Giemsa Staining For staining blood
Cytologic Procedures
Immunofluorescent For cancer, enzymes
Specimen smear preparation
and hormones
• Makes use of an adhesive
Fixation Principle involved is
• Preservation of cell integrity antigen-antibody
• To prevent distortion and destruction of reaction
specimen Fluorescent dyes
Requirements for fixation of specimen 1. Rhodamine dye- red
1. Fixative of choice must be previously 2. Fluorescein- variety of yellow
prepared 3. Acridine- pinkish
2. Place in a couplin jar with cover or Mounting
container with cover to prevent • Placing of cover glass on the slide plus
evaporation the use of a medium for permanent
3. Volume requirement is 1:1 that means, 1 adhesion
part of specimen to 1 part of fixative Types of Mountant
Choice of fixative for cytology Temp or Wet Uses glycerin-jelly
1. Ether-alcohol mixture
Pern or Dry Uses Canada Balsam
o The primary reagent is alcohol
o The secondary reagents are
Labelling
ether, acetone, chloroforms,
glacial acetic acid
Reading and Interpretation
2. 95% ethyl alcohol
• Scanning of smears and determine
3. Carnoy’s fluid
percentage of normal and abnormal
o Contains absolute alcohol,
cells
chloroform and glacial acetic
acid
Examples of fixatives for such specimen Pap’s Cytologic Report: (Report for Cytologic
1. 50% alcohol Smears)
o For pleural and peritoneal fluid Class 1 Normal cytology
2. 70% alcohol Absence of abnormal cells and if
o For sputum present it only ranges from 0-25%
3. 95% alcohol Class 2 Atypical cytology but no evidence
o For urine, gastric aspirates, CSF of malignancy
Staining Atypical cells range from 26-29%
• Reaction between dye and the cell to Class 3 Doubtful cancer
produce the desired color Cells are either atypical or cancer
Other staining methods cells
Modification of H&E Shorter procedure Ranges from (+) or (-) 50%
than Pap’s stain Class 4 True or definite cancer
Polychrome Ranges from 51-75%
Methylene Blue Class 5 Metastatic cancer
Stain Plenty of cancer cells that ranges
Neef and Pundell For identification of from 76-100%
microorganisms in
smear (like in vaginal Clinical Reports
smear and sputum) Stage 1 Organ
Cresyl Violet Method Stage 2 Surrounding tissue
Periodic Acid Schiff Stage 3 Blood vessel and lymphatic
Staining (PAS) vessels

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Stage 4 Spread to distant organ

Procedures used in Cytologic examination

Old Procedure
1. Simple microscopy
a) Interference microscopy
b) Phase contrast microscopy
c) Dark field microscopy
o Identify certain organism
d) Cytochemistry
o Study of important elements
e) Electron Microscopy
o Identify details of cells
f) X-ray
o Uses gamma radiation
Recent Procedure
RAS (Radio Active Scan)
• Ex: brain scan, liver scan, bone scan
UST (Ultrasound Technique)
• For reproductive organ
RIA (Radio Immuno Assay)
• For enzymes, hormones, drugs
IF (Immunofluorenscence)
• Uses special stain

Histopathologic Technique

• Deals with the preparation of solid tissue

coming from different specimen from a
living or dead body for microscopic
examination

Sources of specimen

Surgical specimen

• Specimen obtained after biopsy or

operation from live person

Autopsy specimen

• Specimen obtained from dead bodies after

post-mortem examination

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