Bladder Wash

Principle:

Cytologic studies are performed on Bladder Wash sediment to detect bladder cancer that may be undetectable by cystoscopy

Collection and Submission:

Bladder washings are obtained by cystoscopy and should be placed in equal parts CytoLyt solution.  The specimen must be labeled with the full patient’s name, second identifier (DOB or MR #), and source.  An electronic order is placed or a Non-Gyn Cytology Requisition completed.  The specimen is sent to Dahl-Chase Diagnostic Services with a completed Non-Gyn Cytology Requisition Form or printed paper copy of the electronic order noting any pertinent clinical information such as history of malignancy. 

Body Fluids

Principle:

Cytology studies are performed on Body Fluids (Pleural, Pericardial, Abdominal, Cul-de-sac, Pelvic washes, and Synovial fluids) to detect metastatic carcinoma, mesothelioma, rheumatoid disease, or inflammatory conditions.

Collection and Submission:

Pleural, pericardial, abdominal, and Synovial fluids are collected by needle aspiration.

Submit fresh and refrigerated along with an aliquot (agitate before decanting) in equal parts 10% 
neutral buffered formalin for cell block.

• Pleural and abdominal (75-250 ml optimal), or pericardial (65-250 ml) fluids: Submit fresh in 2-3 capped 60 cc syringes or sterile containers and refrigerate, and one container with 50:50 mix with 10% neutral buffered formalin for cell block. If less than 60 ml, submit fluid fresh and refrigerated.
• Place electronic order or complete Non-Gyn cytology requisition.
• Label all containers with the patient’s full name, second identifier (DOB or MR#), and source.
• Any remaining fluid should be kept refrigerated at the hospital laboratory for up to 3 days.

Cul-de-sac fluids and abdominal washes are usually collected at laparoscopy. If the specimen has been obtained using saline, submit fresh and refrigerated.

Heparin may be added (5 units/ml of body fluid) if the specimen is clotting. If a small amount of fluid (<5ml) is collected or there will be a delay in transporting, the fluid should be added to an equal amount of Cytolyt solution.

The specimen must be labeled with the patient’s full name, second identifier (DOB or MR #), and source and submitted to Dahl-Chase Diagnostic Services accompanied by a completed Non-Gyn Cytology Request Form or printed paper copy of electronic order noting any pertinent clinical information such as history of malignancy.

Bronchoscopy Specimens

Principle:

Cytology studies are performed on bronchoscopy specimens (bronchial brush, bronchial trap, and bronchial wash) to detect lung cancer. Herpes virus, Cytomegalovirus (CMV), Aspergillus and Pneumocystis among others may also be detected. Cytology studies are performed on broncho alveolar lavage to detect vegetable cells, microorganisms, ferruginous bodies, lipid-laden macrophages, and hemosiderin-laden macrophages. Cytology studies are performed on Wang needle aspirations of the trachea to assess mediastinal disease.

Collection and Submission:

Bronchial brushings are obtained from a specific area of the bronchial tree during bronchoscopy. The brush from the procedure should be rapidly rotated on the surface of a slide and the smear fixed immediately with 95% alcohol or spray fixative. The final brushing may be rinsed in CytoLyt but cannot be reused in the patient.

Bronchial washings are obtained from a specific area of the bronchial tree by instilling 10ml of normal saline in small portions and re-aspirating the material with suction. The material collected should be submitted fresh and refrigerated.

Bronchial traps are the mucus traps that are used to suction secretions during the bronchoscopy procedure. Material from any area of the lung that was sampled would be present in the trap, which should be submitted fresh and refrigerated.

Broncho alveolar lavage specimens are obtained when the bronchoscope is advanced into a bronchial segment until it gently occludes the lumen. Aliquots of 20-60 ml of sterile normal saline are infused and re-aspirated and collected in a suction trap. If this is part of a bronchoscopy procedure, the specimen may be submitted with the other material collected fresh and refrigerated.

Wang from bronchoscopy is obtained by needle aspirations through the trachea wall into the mediastinum with the use of a Wang needle. The specimen is aspirated into the needle and the needle is withdrawn and rinsed in CytoLyt solution. Alternatively, optional smears (maximum of 2) may be made by smearing the aspirated material between two glass slides and immediately fix in 95% alcohol or spray fixative.

All specimens and slides must be labeled with patient’s full name, second identifier (DOB or MR #), and source and submitted to Dahl-Chase Diagnostic Services with a completed Non-Gyn Requisition or printed electronic order noting any pertinent clinical information. All the specimens that are collected may be submitted with one requisition or electronic order.

Specimens for Special Stains

Special stains may be performed to highlight the presence of microorganisms, lipid, and iron. Special stain slides are made in addition to the routine cytology material. Broncho alveolar lavage is the preferred specimen for all stains, but they may be performed on bronchial washings, bronchial traps, and bronchial brushes. Sputum is less than optimal for special stains.

Collection and Submission

• GMS-pneumocystis for the detection of Pneumocystis
• GMS-fungus for the detection of fungus including Candida and Aspergillus
• AFB for the detection of mycobacteria
• Iron stain for the detection of hemosiderin-laden macrophages

These stains are performed when ordered on the requisition or electronic order.

• Fat stain for detection of lipid laden macrophages. A fresh specimen (unfixed) must be submitted for detection of lipids in specimens. The fresh specimen must be submitted in addition to the routine cytology material.

CSF and Vitreous Fluid

Principle: 

Cytologic studies are performed on CSF and Vitreous fluid to detect metastatic carcinoma, lymphoma, leukemia, cryptococcus, and inflammation.

Collection and Submission:

CSF is collected at lumbar puncture. At least 1ml of CSF should be received fresh to Dahl-Chase within 1 hour of collection. This specimen should be sent without delay, especially for suspicion of leukemia and lymphoma. If the specimen is not received within 1 hour of collection, it must be refrigerated. If the delay in receipt is more than 72 hours, it must be fixed with an equal amount of CytoLyt. The specimen must be labeled with the patient’s full name, second identifier, (DOB or MR #), and source and submitted to Dahl-Chase Diagnostic Services accompanied by a completed Non-Gyn Cytology Requisition or printed paper copy of electronic order noting any pertinent clinical information such as history of malignancy or lymphoma.

Vitreous fluid is obtained by aspirate or washing. At least 1ml of vitreous fluid should be sent fresh, refrigerated to Dahl-Chase. The specimen must be labeled with the patient’s full name, second identifier (DOB or MR#), and source and submitted to Dahl-Chase Diagnostic Services accompanied by a completed Non-Gyn Cytology Requisition Form or printed paper copy of electronic order, noting any pertinent clinical information.

Specimens for special stains

GMS-Fungus for detection of fungus including Cryptococcus.

Gastrointestinal Tract Brushings

Principle: 

Cytologic studies are performed on the gastrointestinal tract to detect or rule out cancer.  Esophageal brushings may also detect Herpes or Candida. 

Collection and Submission:

Specimens from the GI tract are obtained by endoscopy. A brushing from a visible lesion is obtained prior to the biopsy. The brush should be rapidly rotated on the surface of the slide and the smear fixed immediately in 95% ethanol or spray fixative to avoid air-drying of the specimen. The slides must be labeled with the patient’s full name, second identifier (DOB or MR #), and source on the frosted end with a #2 lead pencil. The slides are submitted to Dahl-Chase Diagnostic Services accompanied by a completed Non-Gyn Cytology Requisition or printed paper copy of electronic order noting any pertinent clinical information.

Nipple Discharge

Principle: 

Cytology studies are performed on nipple secretions to detect breast carcinoma, or papilloma.

Collection and Submission:

Gently strip subareolar area and nipple with thumb and forefinger.  When secretion occurs, allow a pea-sized drop to accumulate.  Apply a glass slide directly to the nipple and secretion.  Fix the slide immediately with spray fixative or 95% ethanol to avoid air-drying.  The slide must be labeled with the patient’s full name, second identifier (DOB or MR #), and source and submitted to Dahl-Chase Diagnostic Services accompanied by a completed Non-Gyn Cytology Requisition Form 
or printed paper copy of electronic order noting any pertinent clinical information. 

Sputum

Principle:

Cytologic studies are performed on sputum specimens to detect lung cancer and respiratory infections.

Collection and Submission:

The patient should expectorate early morning sputum directly into sterile container which is submitted fresh and refrigerated. Morning specimens resulting from overnight accumulation of secretions yield the best diagnostic results. Three specimens on three successive days should be collected to ensure a maximum of diagnostic accuracy. The sputum specimen must be labeled with the patient’s full name, second identifier (DOB or MR#), and source and date of collection and submitted to Dahl-Chase Diagnostic Services with completed Non-Gyn Cytology Requisition or printed paper copy of electronic order noting any pertinent clinical information.

Tzanck Prep for Herpes

Principle:

Cytologic studies are performed on Tzanck preparations to detect the presence of cells that are infected with Herpes virus.

Collections and Submission:

A suspected lesion should have an intact vesicle to be sampled. Open the vesicle, discarding the top. Fluid in the vesicle may be used for viral culture, otherwise discard the fluid. Scrape the base of the vesicle with a scalpel blade. Try to avoid bleeding. Smear the material collected on the scalpel blade onto a slide and immediately fix the specimen with spray fixative or 95% ethanol before air-drying occurs. The slides must be labeled with the patient’s full name, second identifier (DOB or MR #), and source and submitted to Dahl-Chase Diagnostic Services with completed Non-Gyn Cytology Requisition or printed paper copy of electronic order noting any pertinent clinical information. 

Urine

Principle:

Cytology studies are performed on urine sediment to detect bladder cancer. CMV may also be detected.

Collection and Submission: 

Collect random voided urine at minimum 30 mls (not first morning) in a clean, wide-mouth jar and add equal amounts of urine to Cytolyt solution.

• Fresh, unfixed urine specimens must be received in the lab the same day as collected and must be kept refrigerated.
• Refrigerated urine specimens must be fixed with CytoLyt within 6 hours of collection.

For newborns, add the small amount of urine collected to the same container of CytoLyt solution over an 8-16 hour period.

Because urine specimens may be obtained with a catheter, it is extremely important to note the method of collection as either voided or catheterized on the requisition and the container.
 
The specimen must be labeled with the patient’s full name, second identifier (DOB or MR #), and source and date of collection and submitted to Dahl-Chase Diagnostic Services accompanied by a completed Non-Gyn Requisition or printed paper copy of electronic order noting any pertinent clinical information.

Guidelines for Shipping Cytology Specimens

1. Pap smear slides or Non-Gyn slides must be submitted in cardboard holders that are rigid enough to prevent slide breakage.
2. All slides must be labeled on the frosted end in #2 pencil with the patient’s full name, second identifier (DOB or MR#), and source. Labeling the slide holder is not sufficient as each slide must be labeled.
3. Non-Gyn specimens must be submitted in leak-proof containers. The containers of CytoLyt solution that are provided for specimen fixation and transport are leak-proof.
4. All specimens must additionally be placed in a zip-lock biohazard bag before being sent to Dahl-Chase Diagnostic Services.
5. Do not submit large volume fluids in any large containers such as large glass vacuum bottles or pleuravac containers. Shipping large containers like these can be hazardous to handle.

Guidelines for Preservation

1. Specimens with high mucus content, such as sputum and bronchoscopy specimens, may be held for 24-48 hours without fixative if they are refrigerated. Mucus coats cells, protecting them from degeneration.
2. Specimens with high protein content, such as body fluids, may be held for up to 72 hours without fixation if they are refrigerated. The protein-rich fluid in which the cells are bathed acts as a tissue culture medium, preserving cellular morphology.
3. Specimens with low mucus or protein content, such as urine, must be fixed immediately. Even a 1–2-hour delay will cause cellular degeneration because the fluid medium in which the cells are bathed contains enzymatic agents capable of causing cell degeneration. Fresh, unfixed specimens with suspicion of leukemia or lymphoma must be submitted without delay.
4. Specimens with low pH, such as gastric material, must be fixed within minutes of collection to prevent cellular destruction by hydrochloric acid.
5. Smears made for cytology must be fixed immediately.

All needed materials may be ordered through Dahl-Chase Purchasing 941-8202 or 1-800-464-2332.