Skip to main content
Close

Diagnostic Cytology

Diagnostic cytology is the process of studying cells to identify diseases.  Diagnostic cytology may be used to help identify various types of cancer and certain infections.

Anal specimens

  • To obtain an anal sampling, a Dacron swab or cytobrush is inserted approximately 1.5 to 2 inches into the anal canal. It is important not to use a cotton swab, as cells tend to cling to cotton and do not release easily into cytology collection fluids.
  • Moisten the Dacron swab with water, not lubricant. Once inserted deep enough into the anus (necessary in order to collect both rectal columnar and anal squamous cells) the swab should be pulled out, applying some pressure to the wall of the anus, rotating the swab in a spiral motion along the way.
  • The cells collected can be placed in a cytology collection fluid (preferred) or spread on a glass slide and immediately spray-fixed.
  • The preferred method of collection is in PreservCytTM collection fluid. The collection device should be thoroughly rinsed and swirled in the vial. 
  • The vial or slide should be labeled with the patient's first and last name, PHN and date of birth. 
  • Place in a specimen bag and ship to the Vancouver Cancer Centre Cytology Laboratory. The specimen must be accompanied by a "PHSA Laboratories Request For Diagnostic Cytology" form with the requested test marked.
 

  • Dacron swab or cytobrush
  • PreservCytTM collection fluid or spray fixative.

 
The "anal pap" is a screening tool used in at-risk populations to identify individuals who have premalignant cytologic changes in their anal epithelium.
The incidence of anal cancer in the general population is less than one case per 100,000. However, when evaluating specific high-risk populations, such as women with cervical lesions and cervical and vulvar cancers, men who are HIV negative with high risk behaviors, and men or women who are HIV positive, the rate is as high as 70 cases per 100,000. Several biological similarities are shared between cervical and anal cancers, including an association with Human Papilloma Virus (HPV) infections.
Anal cytology is suggested as a screening test for selected patients at high-risk for anal squamous intraepithelial lesions (ASIL).
There are no official guidelines regarding anal cytology screening for ASIL. The following patient group information is based on the approach used by the Palefsky group at University of California at San Francisco which is spearheading the clinical research on anal dysplasia (Palefsky 2001).
HIV-negative men or women with a history of receptive anal intercourse or anal warts.
HIV-positive men with a history of anal intercourse or anal warts. Some clinicians screen patients with CD4 counts that are less than 500/mm3 more frequently.
HIV-negative women with a history of anal warts, high-grade cervical squamous intraepithelial lesions (SIL)/carcinoma, or vulvar SIL/carcinoma.
HIV-positive women. Some clinicians screen patients with CD4 counts that are less than 500/mm3 more frequently.
Consider screening patients with organ transplants on chronic immunosuppressive agents.
Anal squamous intraepithelial lesions (ASIL) may be detected in the anal canal and most likely represent the precursor to anal cancer.
ASILs range from low to high grade. High-grade squamous intraepithelial lesions (HSIL) most likely represent true invasive cancer precursor lesions in the cervix, and most likely in the anus. Atypical squamous cells of undetermined significance (ASCUS) may also be found on cytologic examination in both the cervix and the anus, and these lesions are often accompanied by biopsy-proven SIL.
Anoscopic and histologic assessment of anal lesions is critical to classify the lesions accurately, since the grade of anal cytology often does not correspond to that of histology, which remains the gold standard. Still, anal cytology appears to play an invaluable role in detecting and treating high-grade dysplastic lesions before they progress to anal cancer. Any cytologic abnormality should be followed up with high resolution anoscopy and any lesion should be biopsied to confirm the grade of dysplasia.
Anal cytology reports will generally follow the format for cervical cytology. The absence of columnar cells in the smear does not reflect the validity of the sample. The sensitivity, specificity, and predictive value do not hinge on the presence or absence of columnar cells. Some sources, however, recommend that both squamous and columnar cells should be present in samples for adequate interpretation of slides.
Specimen Required: The methodology for this assay is routine cytopathologic evaluation using either conventional smears or the ThinPrep® method.


 

Bile drainage

  • Specimen Required: 10 mL or more of collected bile drainage.
  • Using appropriate sterile technique, collect as much bile drainage through the drainage apparatus as possible, into a clean plastic specimen container with an equal volume of fixative. 
  • Label the container with the patient's first and last name, date of birth, and specimen source. 
  • Submit the specimen and the completed PHSA Laboratories Request For Diagnostic Cytology form to the Vancouver Cancer Centre Cytopathology Laboratory.
 
  • Standard transcutaneous or endoscopic biliary drainage equipment. 
  • Clean plastic specimen container of an appropriate size. 
  • Fixative: Saccomanno or 50% ethyl or methyl alcohol.
 

For the detection of malignant cells arising within the hepatobiliary system

 

Body cavity fluids

  • Using standard paracentesis technique, obtain a fluid specimen from the desired body cavity. If necessary, move the patient into multiple positions to suspend cellular material in the fluid. 
  • A minimum of 50 mL of specimen is desirable for optimal cytologic evaluation. If other studies are required, withdraw a fraction of the specimen and submit it to the appropriate separately, following their guidelines for specimen collection.
  • Heparin should be added to bloody fluids to reduce clotting. Place 3 units of heparin per mL capacity of the collection container.
  • Agitate the container to coat the sides with heparin. Rinse the paracentesis instrument with a small amount of heparin to prevent clotting of specimen before it is put into the collection container. Add specimen to the heparinized container. Gently agitate to thoroughly mix the specimen and heparin.
  • Submit the specimen to the Vancouver Cancer Centre Cytology Laboratory along with the completed PHSA Request For Diagnostic Cytology test form. If transport of the specimen will be delayed more than 24 hours, add an equal volume of 10% buffered formalin (if sample size is too large to accommodate this volume, a well mixed aliquot of the specimen with an equal volume of fixative may be utilized). If transport time will be less than 24 hours, or fixative is not available, the specimen should be refrigerated or kept on wet ice until transport to the lab.
  • Note: recent changes to lab protocol demands the use of 10% buffered formalin as the fixative of choice; please do not use 50% ethyl or methyl alcohol.
 
  • Standard paracentesis equipment. Clean collection container of appropriate size. 
  • Fixative (10% buffered formalin). Optional: heparin for bloody fluids
 

Detection and characterization of malignant cells in body cavity fluids.

Body cavity fluids are serous fluids that accumulate in the body cavities due to a disease process. They are commonly evaluated for the presence of malignant cells from metastatic disease. Body cavity fluids in general are relatively easy to obtain and are relatively difficult to compromise. However, in some instances, due to a large number of inflammatory cells, specimens may degenerate rapidly. In addition, if large amounts of protein are present, the specimen may clot, trapping diagnostic cells within the clot. These fluids include pleural, peritoneal, pericardial, synovial, and pelvic washing.

 

Breast nipple secretion

  • Label the two slides with the patient's first and last name, date of birth and specimen site in pencil on the frosted end. 
  • Collect a small amount of nipple secretion directly onto one of the slides. Oppose a second glass slide onto the first, allowing the collected material to provide surface tension between the two slides, and then gently and quickly pull the two slides apart in a horizontal motion to distribute the material in a thin film over both slides. 
  • The smears may be immediately fixed either by spray fixative or immersion in 95% ethyl alcohol for a minimum of 5 minutes, or allowed to air dry without fixative. Whether fixed or air dried, slides must be completely dry before packaging.
  • Submit the specimen and the completed PHSA Request For Diagnostic form to the Vancouver Cancer Centre Cytology Laboratory. Indicate on the form as to whether the specimen was fixed or air-dried.
 
Two clean glass slides (single-end frosted).
Optional: Spray fixative or 95% ethyl or methyl alcohol


 

Detection of atypical or malignant cells in nipple discharge specimens.

 

Bronchial brushing

  • Using standard bronchoscopy technique, identify the lesion in question and obtain a brushing sample of the lesion. Upon withdrawing the brush, remove sheath, agitate the brush vigorously in 15 ml centrifuge tube(s) with approximately 10 ml of Cytolyt fixative. DO NOT APPLY THE BRUSH DIRECTLY TO SLIDES. If possible, detach the brush and leave it in the vial. 
  • Label the vial with patient's first and last name, date of birth, and specimen source. 
  • Submit the specimen along with the completed PHSA Request For Diagnostic Cytology form to the Vancouver Centre Cytopathology Laboratory.
 

  • Standard bronchoscopy equipment.
  • One (or more if necessary) 15 ml centrifuge tube(s) with approximately 10 ml of Cytolyt fixative.

 
For the detection and characterization of bronchoscopically visible premalignant/malignant pulmonary lesions; for the identification of some microbiologic pathogens (primarily viral and fungal).
Specimen Required: Bronchoscopically-directed brushing of the identified lesion.


 

Bronchial washing

  • Specimen Required: Bronchoscopically-obtained washing (at least 10 ml is preferred) of the bronchi in the region of the suspected lesion.
  •  Using standard bronchoscopy technique, lavage the distribution of the bronchus to be sampled. 
  • Collect the wash in a clean container. Add equal volume of fixative. 
  • Label the container with patient's first and last name, date of birth, and specimen source. 
  • Submit the specimen, along with the completed PHSA Request For Diagnostic Cytology form to the Vancouver Cancer Centre
 

Standard bronchoscopy equipment. 120 mL clean plastic specimen container(s). Fixative ( 50% ethyl or methyl alcohol, or CytoLyt (recommended) ).

 

For the detection and characterization of bronchoscopically ill-defined or invisible premalignant/malignant pulmonary lesions; for the identification of some microbiologic pathogens (primarily viral or Pneumocystis carinii).

 

Bronchoalveolar lavage

  • Specimen Required: Bronchoscopically-obtained lavage (preferably at least 20 mL) of the distal airways and alveoli.
  • Using standard bronchoscopy BAL technique, lavage the lung distribution in question with sterile, normal saline (or other physiologic solution). 
  • Collect the lavage specimen in a clean specimen container. 
  • Label the container with the patient's first and last name, PHN, date of birth, and specimen type.
  • Note: The Cytology portion of the BAL should be fixed with an equal volume of 50% ethyl or methyl alcohol or CytoLyt. 
  • Submit the Cytology specimen, along with the completed PHSA Request For Diagnostic Cytology test form, to the Vancouver Cancer Centre Cytology
 
  • Standard bronchoscopy equipment. 
  • 120 mL sterile plastic specimen container, 50 % ethyl or methyl alcohol or CytoLyt.
 

For the detection and characterization of microbiologic pathogens (primarily Pneumocystis carinii, viral, fungal, and bacterial) in immunocompromised patients ; detection and characterization of malignancy.

 

CSF (cerebrospinal fluid)

  • Specimen Required: Minimum 1 ml CSF. 
  • If possible deliver fresh (unfixed) to the lab within 24hrs (refrigerate, ship cool; do not freeze) 
  • Otherwise, preserve by immediately adding an equal volume of 50% ethyl or methyl alcohol or CytoLyt.
  • Express into tube and label the container with the patient's first and last name, PHN, date of birth, and specimen type.
  • If delivering within 24 hours place on ice immediately and forward to BC Cancer Labs. Otherwise add fixative immediately and mix well. 
  • Submit the specimen and the completed PHSA Request For Diagnostic form to the Vancouver Cancer Centre Cytology Laboratory.
 
  • Clean sterile container such as a red top tube. 
  • Fixative: 50% ethyl or methyl alcohol or CytoLyt.
 

Cytologic evaluation of cerebrospinal fluid (CSF) is an effective means for diagnosing many disorders involving the central nervous system (CNS).

 

Fine needle aspirate (FNA)

  • Option 1: From 2 aspirates prepare two separate smears directly onto slides, one air dried and one immediately spray fixed with an alcohol based fixative (Cytofix) or immediately immersed in 95% methyl or ethyl alcohol. 
  • If the slide is directly immersed it should be allowed to fix for a minimum of 2 minutes and then allowed to air dry prior to packaging. Then rinse needles from each aspirate into CytoLyt®.
  • Option 2: Rinse all aspirated material directly into CytoLyt®
 
  • 3, 5, 10, or 20 mL syringe. 
  • Syringe pistol (optional). 
  • 22 to 25 gauge needle of appropriate length. 
  • Single-end frosted glass slides labeled with the patient's first and last name, date of birth, and specimen source (for preparation of direct smears). 
  • Spray fixative or 95% ethyl or methyl alcohol
 

To diagnose mass lesions, and to characterize the type of malignancy or benign disease present.

 

Gastric brushing

  • Specimen Required: Endoscopically-directed brushing sample of the identified lesion.
  • Instruct the patient to fast overnight or for a minimum of six hours prior to the procedure. Using standard endoscopy technique, identify the lesion in question and obtain a brushing sample of the lesion.
  • Note: It is important to brush the edges of an ulcer, as well as the floor, in order to obtain diagnostic material.
  • Upon withdrawing the brush, remove sheath, agitate the brush vigorously in the 15 ml centrifuge tube of Cytolyt fixative. Do not apply the brush directly to slides. If possible, detach the brush and leave it in the 15 ml centrifuge tube. 
  • Label with the patient's first and last name, date of birth, and specimen source. 
  • Submit the specimen along with the completed PHSA Cytology test request form to the Vancouver Cancer Centre Cytopathology Laboratory.
 
  • Standard endoscopy equipment. 
  • One (or more if necessary) 15 ml centrifuge tube(s) with approximately 10 ml of Cytolyt fixative.
 

For detection and characterization of endoscopically visible gastrointestinal lesions; for the identification of some microbiologic pathogens (primarily herpes, CMV, and Candida).

 

Gastric washing

  • Specimen Required: Endoscopically obtained washing (preferably at least 10 mL) of the region of the suspected lesion.   
  • Instruct the patient to fast overnight or for a minimum of six hours prior to the procedure. Using standard endoscopy technique, lavage the area of interest using a physiologic solution. Aspirate the solution and place in a clean specimen container with an equal volume of fixative. 
  • Label the container with the patient's first and last name, date of birth, and specimen source. 
  • Submit the specimen and the completed PHSA Laboratories Request For Diagnostic Cytology form to the Vancouver Cancer Centre Cytopathology Laboratory.
 
  • Standard endoscopy equipment. 
  • 120 mL clean plastic specimen container(s). 
  • Fixative (either Saccomanno fixative or 50% ethyl or methyl alcohol).
 

For detection and characterization of endoscopically ill-defined or invisible gastrointestinal lesions; for the identification of some microbiologic pathogens (primarily herpes, CMV, and Candida).

 

Oral scraping or brushing

  • Specimen Required: Direct smear of material collected from the oral mucosa.
  • Label the slides with the patient's first and last name, date of birth, and specimen source in pencil on the frosted end. 
  • Gently scrape the area of abnormality with spatula. Quickly and evenly smear the collected material on a glass slide.
  • Immediately spray fix or immerse the slide in fixative. Repeat the process with the second slide if necessary for better diagnostic yield. Repeat the process for additional areas if necessary. 
  • Submit the specimen and the completed PHSA Request For Diagnostic Cytology form to the Vancouver Cancer Centre Cytopathology Laboratory.
 
  • One (or more) clean glass slides, 
  • Spray fixative, or 95% ethyl or methyl alcohol, oral scraping spatula.
 

Detection and characterization of malignancy and infectious processes in the oral cavity.

 

Pelvic washing

  • Specimen Required: 10 mL (or more) of fluid obtained from an appropriately performed washing.            
  • Using appropriate sterile technique during intra-abdominal surgery, instill a physiologic solution into the pelvic cavity. Lavage the area of interest. 
  • Aspirate the solution and place in a clean container. 
  • Label the container with the patient's first and last name, PHN, date of birth, specimen type and collection date. 
  • Submit the specimen and completed PHSA Request For Diagnostic Cytology test request form to the Vancouver Cancer Centre Cytology Laboratory. 
  • If transport of the specimen will be delayed more than 24 hours, add an equal volume of 10% buffered formalin. If transport time will be less than 24 hours, or fixative is not available, the specimen should be refrigerated or kept on wet ice until transport to the lab.
 
  • Standard suction equipment. 
  • Clean collection container of appropriate size. Fixative (10% buffered formalin)
 

Detection and characterization of malignant cells in pelvic washing.

 

Sputum

  • Sputum specimens should be obtained as follows when clinically feasible. The optimum time for specimen collection is within 15 to 30 minutes after waking and before eating breakfast. Brushing of teeth or rinsing of the mouth thoroughly with water will reduce contamination by saliva. 
  • Instruct the patient to inhale and exhale deeply, forcing air from the lungs using the diaphragm. Repeat until the patient coughs and is able to produce a sputum specimen. 
  • Collect the specimen in the container attempting to obtain at least one teaspoon of sputum. Specimen should be a deep cough specimen and not saliva. Saliva is of no diagnostic value.
  • Collect the sputum in a clean container. Add equal volume of fixative. 
  • Label the container with patient's first and last name, date of birth, and specimen source. 
  • Submit the specimen, along with the completed PHSA Request For Diagnostic Cytology form to the Vancouver Cancer Centre
 
  • ‎120 mL clean plastic specimen container;
  • fixative (either Cytolyt™ cytology preservative or 50% ethanol or methanol).
 

  • For the detection and characterization of premalignant/malignant pulmonary lesions.
 
Urine (voided or catheterized)

  • Specimen Required: Preferably about 50 ml of an appropriately-collected voided or catheterized urine specimen.
  • For purposes of obtaining the greatest yield of diagnostic material, do not collect the first voided urine of the day. The second voided urine specimen should be obtained, if possible. A midstream, clean-catch specimen is recommended to avoid vaginal contamination in female patients. A midstream specimen, not necessarily clean catch, is recommended for male patients. If the patient must be catheterized to obtain the specimen, this should be noted on the test request form, as catheterization can lead to artifacts which may be otherwise misinterpreted. 
  • Add equal volume of fixative to the specimen. Ideally fixative should be added immediately. If fixative is not available, the specimen should be refrigerated or kept on wet ice until transport to the lab. 
  • Submit the specimen to the PHSA Request For Diagnostic Cytology form to the Vancouver Cancer Centre Cytopathology
 
  • Clean collection container of appropriate size. 
  • Standard catheterization equipment (for catheterized urine). 
  • Fixative (50% ethyl or methyl alcohol).
 

Detection and characterization of malignant cells and other urologic abnormalities in symptomatic (usually hematuria) patients; screening for malignancy in selected individuals at high risk for the development of urologic malignancy; detection and characterization of some nonneoplastic renal diseases in symptomatic (usually hematuria) patients.

  

Urological specimens (other)

  • Specimen Required: 10 mL (or more) of an appropriately-collected, cystoscopically-derived specimen.
  • Washing: Using standard cystoscopy technique, obtain washing specimens, carefully denoting specific specimen sites for each specimen on the test request form.
  • Add an equal volume of fixative to the specimen container. 
  • Submit the specimen to the Vancouver Cancer Cytopathology Laboratory along with the completed PHSA Laboratories Request For Diagnostic Cytology form. If fixative is not available, the specimen should be refrigerated or kept on wet ice until transport to the lab.
  • Brushing: Using standard cystoscopy technique, identify the lesion in question and obtain a brushing sample of the lesion.
  • Note: It is important to brush the edges of an ulcer, as well as the floor, in order to obtain diagnostic material. 
  • Upon withdrawing the brush, agitate the brush vigorously in a 15 ml centrifuge tube with approximately 10 ml of 50% ethyl or methyl alcohol. Do not apply the brush directly to slides. If possible, detach the brush and leave it in the vial. 
  • Label the vial with the patient's first and last name, date of birth and specimen source. 
  • Submit the specimen along with the completed PHSA Laboratories Request For Diagnostic Cytology form to the Vancouver Cancer Centre Cytopathology Laboratory.
 
  • Standard cystoscopy equipment. 
  • Clean collection container of appropriate size. 
  • Fixative (50% ethyl or methyl alcohol).
 

Detection of suspected malignancy utilizing lavage and brushing specimens obtained cystoscopically (bladder washing, renal pelvis washing/brushing, ureteral washing/brushing or urethral washing/brushing); staging of urologic malignancies.

 

SOURCE: Diagnostic Cytology ( )
Page printed: . Unofficial document if printed. Please refer to SOURCE for latest information.

Copyright © BC Cancer Agency. All Rights Reserved.

    Copyright © 2017 Provincial Health Services Authority