Unit 10 Flashcards
- Peritoneal, pericardial and pleural fluids;
- CSF;
- Nipple discharge;
- Bronchial brushings/washings;
- Sputum;
- *Gastric washings;
- Urine sediment;
- Prostatic secretions/fluid;
- Cervicovaginal (paps) smear
SPECIMEN FOR EXAMINATION
● Volume;
○ Measure using the graduated cylinder or syringe (if the volume is small).
○ mL is used.
● Color;
● Consistency.
○ To check whether the specimen is watery,
mucoid, purulent, etc.
LIQUID FORM
● Number of slides received;
○ Usually in a fixed state.
○ Document how many slides are submitted
which are part of the documentation.
● Labeled specimen.
PREPARED SMEARS
● Urinary Sediment;
● Bronchial Lavage Specimen;
● Specimens that utilize proteolytic enzymes during processing.
○ Examples
■ Trypsin
■ Conc. Sputum
■ Enzymatic lavage sample from the GIT
SPECIMENS THAT REQUIRE ADHESIVE AGENT
The more watery the specimen is, the more difficult the adherence of the specimen to the slide.
TRUE
● Must be permeable to both fixative and the stain;
● Must not retain the stain.
CHARACTERISTICS OF A GOOD ADHESIVE
● Pooled human serum or plasma;
● Celloidin Ether-Alcohol;
● Leukonostoc culture.
GOOD ADHESIVES FOR CYTOLOGIC METHOD
● Smears should be from fresh material;
● Complete requisition data;
○ Patient’s ID: name, age, gender, LMP, PMP,
date, and type of specimen
● Label the slide.
SMEAR PREPARATION
- Streaking;
- Spreading;
- Pull apart;
- Touch or impression smear (Imprint).
- All have to be fixed immediately in the fixative of choice.
METHODS OF SMEAR PREPARATION
- Cellblock Preparation
A histologic preparation that is a sediment
that would be processed just like a tissue
and will be used as a complement to the
smears.
CELLBLOCK PREPARATION
Used for preparing mucoid secretions, vaginal secretions/pap smear preparation, sputum, and gastric contents.
Use a spatula, dissecting needle or applicator stick and streak in a zigzag fashion.
A spatula or a stainless wire comb loop needle which is streaked in a zigzag fashion.
STREAKING
Used for thick mucoid secretions (smears of fresh sputum and bronchial aspirates).
The material is placed at the center of the slide and gently spread out in a centrifugal fashion where it is thin and evenly distributed.
Star pattern: another spreading technique that may also be done wherein the material is gently teased in a centrifugal fashion forming a star-like pattern.
Twist method can also be done.
SPREADING
We have two slides wherein the material is sandwiched and macerated in between and gently pressed/squashed in opposite directions using both fingers.
PULL-APART
Advantage: Final results consist of two smear slides. The material is more thin and evenly.
Thinner material = preferred = better cytologic evaluation.
pull-apart method
For preparation of direct impression from the cut surface of tissue like the lymph nodes, mucosa, and other surgical /autopsy material.
TOUCH OR IMPRESSION SLIDE
Impression cytology being collected from a patient with herpes simplex keratitis, using a sterile glass slide with polished edges.
A sterile glass slide is used and gently imprints/ touches the cornea surface → fragments are transferred/adhered to the slide.
The touch imprint complements a frozen section.
TOUCH OR IMPRESSION SLIDE
- Using standard paracentesis technique,
obtain a MINIMUM 10ML fluid specimen from
body cavity. HEPARIN may be added to the
specimen to reduce clotting. - Place three (3) units of heparin per mL
capacity of the collection container. Gently
agitate to thoroughly mix the specimen with
heparin. Submit the specimen along with the
completed cytology request form. - The specimen should be refrigerated or
kept on wet ice until transported to the lab.
● Aside from the liquid specimen, smears can also be used. In most instances, the pull-apart method is the most popular and easiest technique to do.
● For specimens that will be delayed: all liquid
specimens must kept in the refrigerator to avoid the generation of the cytologic material.
○ Exception: CSF–considered as stab material,
meaning should be collected and processed
immediately.
COLLECTION OF SPECIMEN: BODY FLUIDS
Fluid that accumulates in the lungs and is aspirated using the aspiration technique.
The liquid material collected is called the pleural fluid/pleural effusion.
The fluid can be smeared onto the slide using the pull-apart method or cell block technique.
PLEURAL FLUID: THORACENTESIS
Another method of collecting specimens that are abnormally seen/accumulates in the peritoneal cavity.
Fluid is referred to as Ascitic fluid/Peritoneal fluid
ASCITIC FLUID: PARACENTESIS
Fluid that is found on the pericardial sac/pericardium.
Fluid can be directly placed onto the slide and processed to form a cell block.
PERICARDIAL EFFUSION: PERICARDIOCENTESIS
L3 and L4 of the spinal column
Manner of collection: Lumbar tap/puncture.
Should be collected and processed immediately.
CSF: LUMBAR TAP
Help diagnose breast cancer early in the disease process.
Used to aspirate material from palpable masses as seen in the image.
Used to diagnose breast cancer in the early phase of its development.
A hypodermic syringe is used, and the fine needle gauge is between 22-25.
The aspirated material is gently placed onto a slide and a smear is prepared.
FINE NEEDLE ASPIRATION BIOPSY (FNAB)
- Gently strip the sub-areolar area and nipple with the thumb and forefinger.
- Place the slide upon the nipple and draw it quickly across the nipple.
-> If 1 drop is obtained, get another slide and do the pull-apart technique. - Immediately immerse the slide into a bottle of 95% Ethyl or Isopropyl ROH or use a spray fixative.
- Label the secretions obtained indicating laterality (left or right breast)
Advantage: Immunohistochemistry can be done directly from the sample as seen in D.
BREAST: NIPPLE DISCHARGE - TOUCH (IMPRINT CYTOLOGY)
Bronchial aspirate;
Bronchial brushings;
Bronchial lavage (BAL).
BRONCHOSCOPY
Specimen: Bronchoscopically-directed brushing of the identified lesion.
Collection Procedure:
1. Using standard bronchoscopy technique, identify the lesion in question and obtain a brushing sample of the lesion.
2. Gently apply the sample on to glass slide and immediately immerse slide into 3% GLACIAL ACETIC ACID ALCOHOL FIXATIVE
3. The brush tip should also be cut off into the solution.
BRONCHIAL BRUSHINGS
Specimen: Bronchoscopically-obtained washing (preferable at least 10 mL) of the bronchi in the region of the suspected lesion.
Collection Procedure:
1. Using standard bronchoscopy technique, lavage the distribution of the bronchus to be sampled.
2. Collect the wash in a clean container. Label the container with correct patient information.
BRONCHIAL WASHINGS
Obtained at three consecutive morning sputum specimens by deep cough method.
Collect the sample in a wide-mouth container containing Saccomano fluid (50% EtOH and 2% carbowax)
SPUTUM CYTOLOGY: NORMAL
Inhalation of aerosol solution for 20 mins to produce a deep cough sample.
Collect the sample in a wide-mouth container containing Saccomano fluid (50% EtOH and 2% carbowax)
SPUTUM CYTOLOGY: INDUCED
Exfoliated cells decompose/autolyze rapidly (whether mechanically or physiologically from the body), which may destroy cellular and nuclear details and will give inadequate results for diagnosis.
Why should FIXATION be done immediately?
All are liquid fixatives = Wet fixation
- Equal parts of 95% EtOh and Ether
-> Ether: highly flammable and ignite spontaneously, also a mild anesthetic (continued inhalation may cause drowsiness). - 95% EtOH
- Carnoy’s fluid
- Equal parts of tertiary butyl alcohol and 1 part 95% EtOH
- SCHAUDINN’S FLUID – sat. aq. Hg2Cl, absolute HoAc
- Methanol – for dried films, bone marrow smears, blood smears, etc.
- Conventional hairspray
Common Fixatives
A cell block is a method of preparing cytologic material so that it can be processed, sectioned, stained, and viewed as a histology section.
Pass through the paraffin processing technique
CELL BLOCK
It can provide diagnostic information in addition to that obtained from cytology slides.
Advantage: complements the smears.
CELL BLOCK
It is easier to do special stains, and immunohistochemistry on cell block slides rather than on regular cytology slides because the slides often require adaptations of the staining protocols and different controls.
TRUE
‘_____’ in this context is not a reference to placing the cells in a paraffin block but refers to the technique of ‘______’ something to hold it together and maintain its shape.
Block; Blocking
Cell blocks are micro biopsies embedded in paraffin that broaden the diagnostic value of cytology specimens and are complementary to cytology preparations.
Cell block is fixed in a 10% neutral buffered formalin.
It employs retrieval of small tissue fragments from FNA specimens which are processed to form a paraffin block.
TRUE
The supernatant is decanted and the sediment is directly placed on the slide and uses proper methods such as pull-apart or streaking.
Plasma Thrombin Method: can also be used wherein the plasma + thrombin is added until the preparation forms a semi-solid clot fixed in formalin.
Another option: Adding agar to the sediment and is fixed with formalin and embedded in paraffin, hence, cell block.
CELL BLOCK TECHNIQUE
