Filling in a request form, glass slides, and object cases
Request form
Cytodiagnosis is a morphological test that utilizes a wide variety of clinical information for reference.
To put in your request for cytodiagnosis, please provide the following specifics in the cytodiagnosis request form.
Please make sure that no information is missing as the result of cytodiagnosis is determined by these factors.
a) Subject's name, sex, age, date of birth, department, etc.
b) Testing materials and methods of their collection
c) Clinical diagnosis, clinical course, testing, treatments, etc.
d) Size and characteristics of lesion
e) For gynecological or mammary gland samples: menstruation and other relevant considerations
f) Previous sample registration No.
g) Number of glass slides provided
Glass slides
a) Write the subject name on the opaque part.
b) For dry-fixed smears, note “ dry.”
Object cases
a) Subject's name, sex, and age
b) Number of sample slides (number of wet-fixed slides and number of dry-fixed slides)
Smear sampling methods vary depending on the type and properties of testing materials. Please refer to the following:
Fixing samples
Fixing is to stop cytological change, such as cytopathy and cytolysis, from occurring.
It is necessary to fix a smear as soon as it is collected.Wet fixation is a technique for Papanicolaou stain and PAS, while dry fixation is used for Giemsa staining.
Whether or not a sample is well fixed is one of the important factors that determine the results of cytodiagnosis, along with the site and method of sample collection;
please ensure that your sample is promptly fixed.
Wet fixation [Gynecologic cytodiagnosis / general cytodiagnosis]
Immerse a smeared glass slide in 95% ethanol for 30 minutes or longer to fix.
Wet fixation methods such as one using CytoTrack™ may also beused.
If using a fixative for coating, make sure that the entire surface of a smear is evenly covered with fixative, either by spraying or instillation.
Dry fixation [General cytodiagnosis]
As soon as a smear has been collected, dry the smear surface quickly using a fan or a hair dryer on the cold shot setting.
Natural drying is not adequate as it may not dry the surface evenly.
■Testing materials and sample volume required
[General cytodiagnosis (00541 0)(0541 8)]
Sputum
Place sputum of a size of red bean between two glass slides, and spread it evenly by rubbing the slides onto one another, moving side to side (cover slip method).
Cancer cells are often found in a larger quantity in blood-strained sputum, or turbid or gel-like viscous parts: collect a smear from these parts.
Sample volume：wet-fixed smear 2 slides
Scrapings (e.g., bronchus, digestive tract, pleura, peritonea)
Scrape a sample off from the focus, etc., using a brush, cotton swab, etc., and smear it onto a glass slide.
It is essential to collect and fix a smear quickly as scrapings dry very fast.
Sample volume：wet-fixed smear 1 slide, dry-fixed smear 1 slide
Fluid materials (e.g., urine, pleural fluid, ascites, pericardial fluid, hydrocele fluid, and lavage fluids)
After centrifuge at 1500 rpm for 5 minutes, smear the sediment either by cover slip method or wedge method.
In the case of a blood sample, smear the buffy coat which contains more nucleated cells.
Sample volume: wet-fixed smear 1 slide and dry-fixed smear 1 slide for urine, pleural fluid, ascites, pericardial fluid, hydrocele fluid, and various lavage fluids
Needle puncture / aspiration samples (e.g., mammary gland, thyroid, lung, lymph nodes, prostatic gland, testis, soft tissue tumor, liver)
Gently eject the needle-aspirated material onto the center of a glass slide to collect smear.
If the sample is in a very small amount, place another glass slide onto the first, then remove it to produce two slides.
If there is a large amount of sample, use wedge method to collect a smear.
Sample volume：wet-fixed smear 1 slide, dry-fixed smear 1 slide
Touch imprint samples (e.g., fixed tumors, lymph nodes)
Gently apply a freshly sliced surface of the tissue that has been cut with a surgeon's knife or blade onto a glass slide.
Sample volume：wet-fixed smear 1 slide, dry-fixed smear 1 slide
Crush preparation (e.g., central nervous system tumor, thyroid tumor)
Place a small mass of tissue (1-2 mm diameter) between two glass slides and press gently to spread the tissue. Remove one slide from the other, and wet-fix the thicker one and dry-fix the thinner.
Sample volume：wet-fixed smear 1 slide, dry-fixed smear 1 slide
＜Note＞
Time allowed from the sample collection to the smear fixing (room temperature)
Sputum, pleural fluid, ascites, pericardial fluid・・・up to 12 hours
Bile, pancreatic fluid, duodenal fluid・・・up to 1 hour, collected in an ice-chilled container
Urine, aspirate, lavage fluid, cerebrospinal fluid・・・up to 1 hour
Scrapings, touch imprint, crush-prepared samples・・・up to 5 seconds
Gynecologic samples are quick to dry: should be wet-fixed immediately upon collecting a smear.
Sample volume：wet-fixed smear 1 slide
Reporting test results
1. The Bethesda system
Determining whether or not a sample is appropriate
Appropriate sample：Well-preserved, with the number of squamous cells that are clearly visible being 8000–12000 with the direct smear, or 5000 or larger with LBC.
Inappropriate sample：Over 75% of the squamous cells are not clearly defined, covered in inflammatory cells or bleeding, or a presence of artifacts due to excessive drying, etc.
＊When a sample is deemed appropriate, cytologic diagnosis is made according to the following classification.
BETHESDA CATEGORY
DEFINITION
ESTIMATED LESIONS
Negative (NILM)
Negative
Organisms Other non-neoplastic findings
Epithelial cell abnortmalities
ASC-US
Atypical squamous cells of undetermined significance
