Exfoliated cytology samples

Sputum:

1.      A sputum series for cytology consists of a fresh, early morning specimen, each day for three days. DO NOT SUBMIT 24-HOUR SPECIMENS. When sputum studies are requested for both cytology and microbiology, two separate specimens must be obtained, each accompanied by the proper request slip. The recommended procedure is as follows:

 

a.      Materials:

i.        Sterile wide mouthed cup with screw top, or;

i.        Hologic 30 mL ThinPrep® Non-Gyn CytoLyt® pre-filled cup. 

 

b.     Collection Method:

i.        Give the patient a clean sputum cup; or, a pre-filled CytoLyt collection cup.

ii.      Have the patient rinse their mouth with water to remove food debris. Instruct the patient to expectorate by a deep cough from the diaphragm and collect the material into the respective collection cup.

iii.    The cup should be delivered to SESP SARC.

iv.    NOTE: PreservCyt media is toxic if swallowed, inhaled, or exposed to skin. Ensure the patient does not rinse their mouth, inhale the vapors, or their skin or mucous membranes to the solution.

 

Oral and oropharyngeal:

1.      Direct scraping prep with glass slides:

a.      Obtain the specimen with a brush (preferred) or tongue depressor by scraping the entire site of the lesion with moderate pressure, to remove the top layer of epidermis without inducing bleeding. Spread the material collected onto a clean glass microscope slide in such a manner that the resulting smear is evenly dispersed in the center of the slide. Immediately spray or dip fix one slide with alcohol, leaving the other slide to air-dry.

b.     For suspected Herpetic lesions, prepare a second smear by re-scraping the remaining site of the lesion to obtain remaining basal or lower epidermal cells which may contain viral inclusions. Repeat spread and fixation steps outlined above.

 

2.      Liquid based Non-Gyn preparation:

 

a.      Materials:

i.        Hologic ThinPrep® 20 or 30 mL Hologic ThinPrep® Non-Gyn CytoLyt®.  pre-filled vial; or,

ii.      Sterile collection container; or,

iii.    Sterile container with saline.

 

b.     Collection Method:

i.        Obtain the specimen with the appropriate collection device. Scrape the lesion, using a rotating motion to collect epidermal layer on brush/spatula. 

i.        Rinse the collection device directly into the ThinPrep Pap Test vial containing preservative fluid.  Cap tightly and shake gently to dislodge cells into solution. Label vial with patient’s name and site of collection (if more than one vial or collection site is obtained).

iii. Do not mix samples from multiple sites in the same vial, use a separate vial and brush for each collection site and label vials with source of specimen.

iv.    Place vial into a specimen transport bag or container.  More than one vial can be submitted in a single bag or container as long as each vial is specifically labeled and each site is identified on the vial and requisition.

 

Bronchoscopic Material (bronchial brush, wash and broncho-alveolar lavage fluid):

1.      If microbiological studies are ordered, the collected specimen/collection device should be placed in a sterile container, with or without sterile saline. A separate specimen should be collected for these studies. If a separate specimen cannot be collected and the specimen is shared, no cytological fixative should be added. The specimen should be sent to SARC/Microbiology first for a sterile aliquot to be removed, then sent to Cytology. Submit a separate requisition for each specimen. Smears prepared on-site must be fixed appropriately and identified with two unique forms of patient identification (ie. Name, MRN).

 

Refer to the UC Davis Health Department of Pathology and Laboratory medicine Laboratory Test Directory for specimen collection requirements in other testing sections.

a.      Materials:

i.        Sterile, wide mouthed cup with screw top; or,

ii.      Hologic 30 mL ThinPrep® Non-Gyn CytoLyt® pre-filled cup.

 

b.     Collection Method:

i.        Bronchial Brushing:

1.      Fluid specimens can be submitted fresh. If the specimen is collected fresh, refrigerate and transport with cold pack or ice.

2.      Bronchial brush specimens should be submitted in Hologic ThinPrep® Non-Gyn CytoLyt®. 1 - 2 inches of wire should be left attached to the brush tip when it is submitted.

3.      Direct smears may be prepared on site by rolling the collection brush onto a glass slide prior to placing the brush head into CytoLyt®. The glass slide should immediately spray, or dip fixed with alcohol. Each slide should be labeled as “fixed” or “AD” for air-dried.

iii.    Bronchial Washing:

1.      Bronchial wash specimens should be submitted fresh.

2.      Specimen should be refrigerated and transported with cold pack or ice.

3.      An aliquot for microbiological testing should be performed prior to cytologic processing.

iv.    Bronchoalveolar lavage (BAL):

1.      Bronchoalveolar lavage (BAL) specimens should be submitted fresh.

2.      Specimen should be refrigerated and transported with cold pack or ice.

 

Effusions: Peritoneal fluid (paracentesis), Pleural fluid (thoracentesis), Pericardial fluid, Ascitic Fluid:

 

a.      Materials:

i.        Sterile collection container.

 

b.     Collection Method:

                                                        i.            Refer to the UC Davis Health Department of Pathology and Laboratory medicine Laboratory Test Directory for specimen collection requirements in other testing sections

                                                      ii.            Effusions should be submitted to the laboratory fresh.

                                                    iii.            The entire specimen should be submitted, not an aliquot of the original.

                                                    iv.            Immediately following the procurement procedure, send the collected specimen to SESP SARC.

Urine:

1.      Clinical indications for urine cytology are: hematuria, follow-up for a patient previously treated for urothelial carcinoma (surveillance), and patient at risk for bladder cancer (screening).

2.      Urine Cytology is not an effective screening tool for asymptomatic individuals.

3.      Tumor Cells: The best specimen of urine for tumor cell detection is a morning specimen obtained after the patient has been up and about (not necessarily the first voiding of the day). If the patient is not ambulatory, submit the morning specimen as soon as possible, accompanied by an appropriate lab slip.

4.      Cytomegalic Inclusion Bodies: Urine specimen should be collected in a cooled container suspended in ICE and delivered immediately to SESP SARC.

 

a.      Materials:

i.        Hologic ThinPrep® 20 mL Non-Gyn PreservCyt® pre-filled vial; or,

ii.      Sterile collection container.

 

b.      Collection Method:

NOTE: Urine specimens may be collected into ThinPrep PreservCyt solution. A 2:1 urine-to-PreservCyt ration is required.

i.        Voided Urine:

-       Voided urine should be collected 3-4 hours after the patient has urinated last. First morning voided urine collection should be avoided.

-       Collection of 25 – 100 mL of urine may be necessary to obtain a specimen that is adequately cellular.

ii.      Catheterized Urine:

-       Catheterized urines are prone to harboring microorganisms from a urinary tract infection. Often, a pooled specimen that has been at room temperature for long periods of time will degrade and could mimic a low-grade neoplasm resulting from the catheter tip scraping off benign tissue. These specimens should be interpreted with these thoughts in mind. 

iii.    Bladder Washing:

-       Bladder washings are typically obtained via a catheter where the bladder is irrigated with pulses of sterile normal saline. This produces a freshly exfoliated sample. Blader washings are often collected prior to bladder biopsy sampling. This method can be more ideal for interpretation than voided urines.

 

Synovial Fluid:

 

a.      Materials:

i.        Sterile collection container.

 

b.     Collection Method:

i.        Refer to the UC Davis Health Department of Pathology and Laboratory medicine Laboratory Test Directory for specimen collection requirements in other testing sections.

ii.      Synovial fluids should be submitted to the laboratory fresh.

iii.    Transport on ice and refrigerate until processing.

iv.    Tissue submitted for crystal analysis must be submitted fresh. Do not submit join fluids for crystal analysis in formalin. Water based fixatives, such as formalin, will destroy crystals.

 

Esophageal, duodenal, pancreatic and ureteral brushings or washings:

 

a.      Materials:

i.        Hologic ThinPrep® 30 mL CytoLyt® cup; or,

ii.      Sterile collection container; or,

iii.    Sterile container with saline.

 

b.     Collection Method:

i.        Refer to the UC Davis Health Department of Pathology and Laboratory medicine Laboratory Test Directory for specimen collection requirements in other testing sections. 

ii.      Fluid specimens can be submitted fresh. If the specimen is collected fresh, refrigerate and transport with a cold pack or ice.

iii.    Brush specimens should be submitted in Hologic ThinPrep® Non-Gyn CytoLyt®. 1 - 2 inches of wire should be left attached to the brush tip when it is submitted.

iv.    Direct smears may be prepared on site by rolling the collection brush onto a glass slide prior to placing the brush head into CytoLyt®. The glass slide should be immediately spray, or dip fixed with alcohol; the other left to air dry. Each slide should be labeled as “fixed” or “AD” for air-dried.

v.      Specimen(s) collected in the above media can be transported at ambient temperature. 

 

Breast Secretions, Skin Scrappings, Oral Scrappings (Tzank smears), Eye (conjunctival) Scrappings:

1.    Obtain sample by scraping lesion deeply with tongue depressor or end of Ayers Cervical Spatula. Do not use a cotton tipped swab to collect material as cellular material is trapped in the fibers and will not adequately transfer to the glass slide. Spread sample on labeled slides, fixed immediately with Spray-Fix. Deliver to SESP SARC. Include Cytology request or EMR order, properly filled out, including site of lesion and clinical information as appropriate. Smears prepared on-site must be identified with two unique forms of identification (ie. Name, MRN).

 

Miscellaneous Fluids:

Bile from T-tubes, spinal fluids, etc., are sent unfixed to SARC, 2P340.

Anal Cytology:

1.      The collection of anal cytology samples for the screening of males/females with high-risk history for anal squamous intraepithelial lesion (ASIL) will be outlined in the following procedure.

2.      Glass microscope slides may be prepared via conventional smear technique and spray/dip-fixed in 95% alcohol. Preferably, specimen should be collected in ThinPrep® PreservCyt® Non-GYN solution and transported to the laboratory within 24 hours of collection.

3.      The presence of lubricant on sample may be considered unsatisfactory for interpretation.

4.      Expired ThinPrep® PreservCyt solution, leaked containers, unlabeled or mislabeled vials will be rejected and require recollection.

 

a.      Materials:

ii.      Hologic ThinPrep® 20 mL Non-Gyn PreservCyt® pre-filled vial. 

iii.    Non-absorbent Dacron® swab.

Note: Dacron® is a registered trademark of Invista.

 

b.     Collection Method:

i.        Moisten a Dacron® swab with sterile water, not lubricant. Do not use a cotton swab. Cells adhere to cotton and are not readily released into the fixative media.

ii.      Insert swab approximately 1.5 – 2.0 inches into the anal canal. Collect adequate sample following published collection techniques.

iii.    Remove swab while applying mild to moderate pressure to the wall of the anus while rotating in a spiral motion sampling the transition zone. 

iv.    Thoroughly swish and swirl the swab or brush as quickly as possible in the ThinPrep® PreservCyt® solution by rotating the device in the solution 10 times while pushing against the PreservCyt® vial wall. Swirl the device vigorously to further release material. Discard the collection device. Do not leave the collection device in the PreservCyt® media. Alternatively, prepare slides by spreading the material on a glass microscope slide and immediately spray/dip fix with 95% alcohol cytologic fixative.

v.      Cap the ThinPrep® PreservCyt® vial securely until a “click” is heard and the torque line on the cap passes the torque line on the vial. Label the vial with the patient’s name, medical record number and date of collection.

vi.    Place the vial and requisition form in a specimen bag for transport to the laboratory.

vii.   Ensure the specimen has been allowed to stand in PreservCyt® solution for at least 15 minutes after collection, prior to processing.