How to fill out the request form, glass slides and object case
Request form
A cytodiagnosis test is a morphological test performed with reference to various clinical information.
When requesting a cytodiagnosis test,
please fill out the following information on the dedicated cytodiagnosis test request form.
Please be sure to fill all the requested information, as it is a factor that influences the cytodiagnosis.
a) Name, sex, age, birth date, department name etc.
b) Test material, collection method
c) Clinical diagnosis, clinical course, tests, and treatment matters
d) Size and properties of lesion
e) In the case of gynecological materials and mammary gland materials, various matters such as menstruation
f) Previously registered specimen number
g) Number of slide glasses submitted
Glass slides
a) Please write your name in the frosted glass section.
b) For dry fixation, please write "dry".
Object case
a) Name, sex, age
b) Number of specimens (Number of wet-fixed and dry-fixed slides)
Method of submitting test material [smear, fixation, test material, specimen volume].
Smears of various materials
The method of smearing differs depending on the type and properties of the test material.
Please smear according to the instructions below.
Specimen fixation
Fixation is the process of stopping changes such as degeneration and lysis of cells,
so it is necessary to fix the cells immediately after smear. Wet fixation is for Papanicolaou staining,
PAS staining, etc. Dry fixation is for Giemsa staining, etc.
The quality of fixation, along with the collection site and method,
is one of the important factors that influences the cytodiagnosis decision. Please fix it quickly.
Wet fixation【Gynecological cytodiagnosis, general cytodiagnosis】
Fix the smeared glass slide by immersing it in 95% ethanol for at least 30 minutes.
For wet fixation, there is also a coating fixation method using Cytotac, etc.
If a coating fixative is used, the fixative should be sprayed or dripped thoroughly over
the entire surface of the smear to avoid unevenness.
Dry fixation【General cytodiagnosis】
Immediately after smearing, dry the smeared surface rapidly with a fan or cold air dryer.
Natural drying is inadequate as it causes uneven drying.
Sputum
Place a "red bean" sized sputum between two glass slides and spread it out evenly,
crushing it back and forth, left and right (pull-apart method).
Since cancer cells are mostly found in the blood sputum, opaque white,
and jelly-like mucus areas, smear these areas.
Specimen volume: Wet-fixed smear specimen 2 slides
Rubbing material (bronchus, digestive tract, pleura/peritoneum, etc.)
Use a brush or cotton swab to rub the lesion and smear it on a glass slide.
The smear should be fixed quickly, as the rubbed material tends to dry out very easily.
Specimen volume: Wet-fixed smear specimen 1 slide, Dry-fixed smear specimen 1 slide
Liquid material (Urine, pleural fluid, ascites, pericardial fluid,
scrotal edema fluid, various washing fluid, etc.)
After centrifugation at 1500 rpm for 5 minutes,
smear the sediment by the pull-apart method or the wedge method. For hematological specimens,
smear the buffy-coated portion of the specimen, which contains a large amount of nucleic components.
Specimen volume: Urine, pleural fluid, ascites, pericardial fluid, scrotal edema fluid,
various washing solutions, etc. Wet-fixed smear sample 1 slide, Dry-fixed smear sample 1 slide
Needle-puncture aspiration material
(Breast, thyroid, lung, lymph node, prostate, testicle, soft part tumor, liver, etc.)
Please blow the aspirated material into the center of the glass slide and smear gently.
If the amount of specimen is very small, peel off the other glass slide after aligning it with
the other glass slide to make two slides.
If a larger amount is obtained, or if the volume is larger, smear the materials using the wedge method.
Specimen volume: Wet-fixed smear specimen 1 slide, Dry-fixed smear specimen 1 slide
Imprinted specimen (various fixed tumors, lymph nodes, etc.)
Please lightly imprint the new tissue split surface, cut with a scalpel or razor, on the glass slide.
Specimen volume: Wet-fixed smear specimen 1 slide, Dry-fixed smear specimen 1 slide
Squash specimen (Central nervous system tumors, thyroid tumors, etc.)
Please place a small piece of tissue (1-2 mm in diameter) between two glass slides,
press it lightly, and when the tissue stretches, remove the glass slide,
fix the thicker one with wet-fix method, and fix the thinner one with dry-fix method.
Specimen volume: Wet-fixed smear specimen 1 slide, Dry-fixed smear specimen 1 slide
＜Precaution＞
Allowable time from specimen collection to smear fixation (room temperature)
Wet fixation should be performed immediately after smear as it tends to dry out easily.
Specimen volume: Wet-fixed smear specimen 1 slide
Please note that the following specimens cannot be requested due to specimen handling.
Formalin-fixed specimen
Frozen specimen
Unrepairable damaged specimens
Following test materials
Tissue, nails, hair, stool, blood, gallstones, stones, bone marrow
Test result reporting
1. Bethesda System
▼Regarding evaluation of suitability of specimens
Suitable specimen: The number of well-preserved,
clearly visible squamous epithelium cells should be 8000-12000 for direct smears and at least 5000 for LBCs.
Unsuitable specimen: This applies if 75% or more of the squamous epithelium cells are obscured,
if they are covered by inflammatory cells, hemorrhage, or if artifacts due to excessive dryness are seen.
* The following cell determination will be performed if the specimen is deemed suitable.
: Atypical cytology but no evidence of malignancy.
Class III
: Cytology suggestive of,but not conclusive for malignancy.
Class IIIa
: Probably benign atypia.
Class IIIb
: Malignancy suspected.
Class IV
: Cytology strongly suggestive of malignancy.
Class V
: Cytology conclusive for malignancy.
* For Class determination, I and II can be read as negative,
III, IIIa, and IIIb as suspicious positive, and IV and V as positive.
3. ABC determination
Classification
Cytological findings
Instructing category
A
No histiocytes in sputum
Materials are unsuitable, retest
B
Only normal epithelial cell
Basal cell proliferation
Low-grade atypical squamous cells
Ciliated columnar epithelium cells
No abnormality now
Next periodic test
C
Moderate atypical squamous cells
Columnar epithelial cells with enlarged nuclei and deep staining
Re-smearing and retest within 6 months
D
High-grade (borderline) atypical squamous cells or cells suspicious for malignancy are present
Immediately conduct a detailed test
E
Malignant tumor cells are present.
Criteria and instruction categories for sputum cytodiagnosis
in lung cancer screening (revised 2016) Clinical
and Pathological Lung Cancer Treatment Regulations, 8th ed.
