Lec 11: Cytopreparatory Technique Flashcards
It is a branch of diagnostic medicine that deals with the study of individual cells spread on a slide and stained properly
Cytology
Advantages of Cytology (5)
- Simple
- cost effective
- quick turn around time
- low complication rate
- high diagnostic accuracy
A branch of pathology that studies and diagnoses diseases by evaluation of cellular changes
Diagnostic Cytology
Or
Cytopathology
The branches of Cytology
- exfoliative cytology
- imprint/abraded Cytology
- aspiration cytology
Branches of Cytology
- It is used in studying cells from specimens that do NOT shed cells spontaneously.
- The microscopic examination of cells that have been desquamated/shed or physically removed from epithelial and mucous membranes.
- Study of cells directly taken from the surfaces of excised or incised specimens by touching or imprinting them with a clean glass slide.
- Aspiration Cytology
- Exfoliative Cytology
- Imprint / Abraded / Impression Cytology
Examples of Exfoliative Cytology
Body Fluids
• sputum
• urine
• pleural & peritoneal fluid
• gastric juice
• CSF
Discharge from:
• vagina, cervix, uterus
• nipple
• sinus
• prostatic
Scraping from:
• buccal mucosa
Example of Impression Cytology
Sentinel Lymph Node Biopsy (SLNB)
The spontaneous disintegration of cells or tissues by autologous enzymes
Autolysis
- elongated sex hormone
- a chromatin mass that lies against the inner aspect of the nuclear membrane in cells of normal female
Barr body
[TRUE or FALSE]
Barr bodies are only found in females. There is NONE for male.
TRUE
It is valuable in diagnosis of lesions of the breast, thyroid, lymph nodes, liver, lung, and soft tissue
Aspiration cytology
Examples of body cavity effusions
- CSF
- pleural, peritoneal, pericardial fluid
- synovial fluid
The Classifications of the types of cytology samples
- according to origin
- according to staining method
What are the Types of Cytology samples according to Origin?
- exfoliative cytology samples
- aspiration cytology samples
What are the Types of Cytology samples according to Staining Methods?
- gynecological specimen
- Non-gynecological specimen
What sample is classified as both exfoliated cytology sample and gynecological specimen?
Cervicovaginal smear
Specimen: CSF
- Source:
- Method of collection
- brain & spinal cord
- Lumbar tap/puncture
Specimen: Ascitic fluid
- Source:
- Method of collection
- Abdominal & pelvic cavity
- Peritoneocentesis / paracentesis
Specimen: Pleural fluid
- Source
- Method of collection
- Pleural cavity (lungs)
- Thoracentesis
Specimen: Pericardial fluid
- Source
- Method of collection
- Pericardial cavity
- Pericardiocentesis
Specimen: Synovial fluid
- Source
- Method of collection
- Synovial cavity (joints)
- Arthrocentesis
The samples should be collected in a ___ container.
Clean, non-sterile, and dry
For aspiration for palpable lesion, what is the volume of the syringe and gauge of the needle used?
- 3 to 5 mL syringe
- gauge 20 to 22 needle
For respiratory tract specimens, the sputum is collected in a wide mouth container which contains ____ (substance).
Saccomanno fluid
Composition of saccomanno fluid
50% ethyl alcohol
+
2% carbowax
For the Macroscopic examination what are the factors/characteristics determined? (5)
- specimen origin
- specimen quantity
- specific gravity
- odor (if present)
- gross characteristics
What are the fixatives for the following specimens:
a. Pleural or peritoneal fluid
b. Urine or bronchial aspirate
c. Specimen requiring centrifugation
a. 50% alcohol
b. 95% ethanol
c. 50% or 95% ethyl alcohol
Preservation of Specimen before Smearing
- Specimens with high mucus content
- Specimens with high CHON content
- Specimen with low mucus/CHON content
- Specimen with low pH
- 12 to 24 hours if refrigerated
- 24 to 48 hours if refrigerated
- 1 to 2 hours even if refrigerated
- • collected on ice
• prepared within minutes of collection
[TRUE or FALSE]
- If too much fluid is obtained, it needs to be centrifuged for 5 minutes
- The centrifuge is required if there is little amount of fluid or the fluid is thick.
- True
- False
» NOT required
Preparing Smears
- This technique is similar to how we make peripheral blood smear or thin smears and parasitology.
- This technique is suggested for mucoid samples and viscous fluids.
- In this technique, a slight pressure is added once the two slides come in contact and then you pull apart.
- Also known as Spread Technique
- Also known as Pull Apart Technique
- Pull-Push technique
- Crush technique
- Crush technique
- Pull-Push technique
- Crush technique
Fixation
- Minimum fixation time
- [TRUE or FALSE] prolonged fixation has no consequences provided container is covered tightly
- 15 minutes
- True
Characteristics of a good cytology fixative (9)
- Inactivates autolytic enzyme activity
- penetrates cell rapidly
- minimizes cell shrinkage
- maintains cell morphology
- replaces cell water
- allows stain permeability
- permits cell adhesion to glass
- kills pathogen
- affords a permanent cellular record
Immersion fixatives (4)
- 95% ethyl alcohol and ether
- 95% ethyl alcohol
- Carnoy’s fluid
- Saccomanno’s fixative
What does Carnoy’s fluid contain?
- absolute alcohol
- Chloroform
- glacial acetic acid
The spray fixatives have a dual purpose. What are they?
- fixes cells
- provides protection
A major technique in cytology that is based on viscosity of the sample.
Direct smear preparation / pick & smear technique
The process wherein a cell button or dot of fluids is processed and sectioned on a rotary microtome using the paraffin embedding technique.
Cell block technique
Uses of Cell Block technique (5)
- architectural evaluation
- categorization of tumors
- special stains & immunohistochemistry
- immunophenotyping
- as archival material for future studies
A major technique in cytology wherein cells are filtered from a fluid through a porous membrane and are subsequently fixed and stained.
Membrane Filter technique
membrane filter technique
- what are the two types of filters?
- What is the membrane made of?
- millipore filter
- nucleopore filter
- cellulose acetate/esters
- nitrate film
- plastic or polycarbonate
It has randomly spaced uniform cylindrical holes or pores which act like a sieve.
Filter
Solutions to fit requirements for Membrane Filter Technique
- Small or scanty sample
- Relatively clear / few cells in large volume
- Wash container with 5 to 10 mL of NSS to increase the volume
- DO NOT PERFORM MFT. Best to centrifuge
Disadvantages of MFT (4)
- overcrowding or clumping of cells
- unwanted staining of filter paper
- distortion of morphology
- difficulties in mounting & storing
A major technique used in cytology that concentrates body fluid using an instrument design to deposit cells onto glass slide
Cytocentrifuge technique (Shandon Cytospin Tech)
Advantages of the Cytocentrifuge technique
- ease of use
- quality of results
- operator safety
- a major technique in cytology that has a great advantage of reaching suspicious sites by means of fine needle
- it is done in almost all palpable superficial lesions
Fine Needle Aspiration Cytology
Fine Needle Aspiration Cytology
- Also known as the triple test of ___ (3)
- Length of needle used
- Gauge of needle
- palpation
- radiologic findings
- cytopathologic analysis
- 1.5 cm
-
Inner organ: gauge 22
Breast: gauge 19
Advantages of Fine Needle Aspiration
- fast and early diagnosis
- less pain, less trauma
- no anesthesia
- accurate
- acceptable by patience and doctors
Fine Needle Aspiration Cytology
- Examples of superficial/palpable masses
- Examples of non palpable masses
- breast
- thryoid
- peripheral mass
- mediastinum
- abdominal organs (liver, pancreas)
- retroperitoneal organs (kidney, adrenal glands, lymph nodes)
STAINS
- it stains DNA, RNA, and acid nucleoproteins
- most precise staining of exfoliated cells
- it stains keratin in vulvar CA
- contains the yellow dye orange G
- a polychromatic mixture
- comprises three dyes
- influences pH of the staining solution
- promote staining with light green at low pH
- Harris hematoxylin
- Orange G6
- Eosin azure 50
- Phosphotungstic acid
