Submitting Pathology Specimens


Pathology Specimens

General Instructions

1. Proper pathologic interpretation requires immediate fixation (in formalin or other appropriate fixative) of all tissue

specimens.

2. Specialized testing, including specimens for cytological evaluation or immunofluorescence may have special

fixation requirements (see specific instructions).

3. Specimen containers must be labeled with patient name, specimen site of origin and collection date.

4. A properly completed requisition form must accompany each specimen.

5. Written procedures for proper specimen handling are available upon request.

6. Please contact our Medical Director or the NHI laboratory manager if you have special research needs.

Specimen Rejection

In order to produce proper results, specimens received by the Pathology Laboratory must meet certain criteria. The

submitting physician or the physician’s representative shall be informed when a specimen is less than optimal, or

unacceptable. Failure to follow the listed guidelines may delay specimen processing and diagnosis.

Cytology Specimens:

General Instructions for Cytology Specimens

1. The container must be properly labeled with the patient’s name, collection date and time and the submitting

physician’s name. A specimen requisition slip should be properly completed and should accompany the specimen

to the laboratory in the outer pocket of a plastic collection bag.

2. The specimen container must be marked with the type of specimen and with the type of fixative used. If special

testing is requested, the request for such testing should be included on the specimen requisition.

3. An aliquot of large volume specimens, e.g., thoracentesis fluid, should be submitted for cytologic evaluation.

Mix the specimen well and transfer about 50 ml. to a separate container and add an equal volume of 50% alcohol

(ethanol, methanol, propanol, or isopropanol are essentially equivalent for fixation purposes) DO NOT

TRANSPORT LARGE CONTAINERS OF UNFIXED SPECIMENS. They represent a significant biohazard,

especially since many such specimens are initially collected in a glass container that may break during transport.

4. If there are questions about handling of a cytology specimen, call Aurora Diagnostics Western Pathology or

Nevada Histology for instructions.

Body Fluids

1. CSF, pleural, pericardial, or ascities fluids are handled by the immediate addition of an equal amount of 50%

ethanol or a substitute (see General Instructions for non-Gyn cytology specimens).

Bronchial Brushings and Washings

• The brush should be rapidly rotated on a small area on the surface of a slide and the smear fixed immediately.

• Fixation can be achieved by either a spray fixative or by immediately immersing the slides in 95% ethanol, 70%

ethanol or a substitute as mentioned above (see General Instructions for non-Gyn cytology specimens).

• DO NOT allow the slides to air-dry prior to fixation as this creates artifacts which interfere with interpretation.

• If fluid fixative is used, separate the slides with paper clips and send the fluid fixative with the slides included to

the laboratory. Alternatively, the smears can be removed from the fixative after 15 minutes and air-dried prior to

shipment to the laboratory.

• Bronchial washes are combined with equal parts of 50% ethanol or a substitute as mentioned above (see General

Instructions for non-Gyn cytology specimens).

Fiberoptic Gastrointestinal Tract Brushings

• The brush should be rapidly rotated on a small area on the surface of a slide and the smear fixed immediately.

• Fixation can be achieved by either a spray fixative or by immediately immersing the slide(s) in 95% ethanol, 70%

ethanol or a substitute as mentioned above (see General Instructions for non-Gyn cytology specimens).

• DO NOT allow the slides to air-dry prior to fixation as this creates artifacts which interfere with interpretation.

• If fluid fixative is used, separate the slides with paper clips and send the fluid fixative with the slides included to

the laboratory. Alternatively, the smears can be removed from the fixative after 15 minutes and air-dried prior to

shipment to the laboratory.

Fine Needle Aspiration Specimens

• FNA specimens are usually obtained by the clinician who prepares the smears.

• The material in the needle is expelled onto a glass slide with care being taken to deposit it as a single drop near

one end of the slide. The needle tip, therefore, should be brought into light contact with the slide and a small

amount of fluid should be expressed. This material is then spread by placing another slide on the first thus applying

flat pressure to the droplet. The slides are then slid apart from each other by opposing longitudinal motion of the

slides. An optimal smear is obtained when the end of the slide on which the droplet is placed is grasped between

the thumb and first finger and the end of the slide toward which the fluid droplet is being spread is supported by the

forth and/or fifth finger(s). The slides must not be separated by vertical motion which produces an uneven smear.

• It is desirable to have multiple sets of slides with some air-dried and some fixed. Fixation can be achieved by

either a spray fixative or by immediately immersing the slides in 95% ethanol, 70% ethanol or a substitute as

mentioned above (see General Instructions for non-Gyn cytology specimens).

• DO NOT allow the slides for fixation to air-dry prior to fixation as this creates artifacts that interfere with

interpretation.

• Separate the slides fixed in liquid with paper clips and send the fluid fixative with the slides included to the

laboratory. Alternatively, the smears can be removed from the fixative after 15 minutes and air-dried prior to

shipment to the laboratory. The air-dried slides require no further treatment prior to delivery to the lab.

• The various slides should be labeled "air" or "fixed" as the laboratory uses a different stain depending on how the

slides were prepared. Air-dried slides are stained with Diff Quik while the fixed slides are stained with Papanicalau.

This combination of stains is adequate for the routine handling of material from most FNA sites.

• The slides must also be labeled with the patient’s name.

Other Cytology Specimens

• Call Aurora Diagnostics Western Pathology or Nevada Histology for information about proper handling.

Sputum

• Use a wide-mouth specimen container. All containers used for submission of cytology specimens must be well-

constructed, rigid plastic with a secure lid to prevent leakage.

• If the patient has recently eaten, have him first rinse his mouth with water.

• A first morning specimen is best since this represents an overnight accumulation of sputum and offers the best

chance of obtaining diagnostic material.

• To obtain the specimen, instruct the patient to cough deeply, raising material from deep within the chest and

expectorate directly into the container. Saliva or material from the back of the throat is NOT satisfactory.

• DO NOT collect 24 hour sputums for cytology. One or several deep coughs is usually sufficient.

• After collection, add an equal volume of 50% methanol or a substitute as mentioned above (see General

Instructions for non-Gyn cytology specimens).

• If a series of three sputums are ordered, it is best to get deep cough specimens in the morning for three days.

Urine

• Urine can be obtained by catheterization or by voiding.

• For cytologic evaluation of the bladder, three morning samples of urine, each of about 50 to 100 ml. obtained on

consecutive days are recommended.

• Special procedures such as retrograde catheterization of the ureter or the renal pelvis can also be used to collect

specimens.

• The method of collection and the site should be specified on the specimen requisition slip.

• The specimen must be immediately fixed by adding an equal volume of 50% ethanol or a substitute as mentioned

above (see General Instructions for non-Gyn cytology specimens).

Bone Marrow

• Specimens may include a core biopsy, clotted aspirate, and prepared smears. The core biopsy should be placed in 10 percent buffered Formalin immediately after touch preps have been made. Fixative may be obtained by calling Nevada Histology.
• Aspirated bone marrow should be collected in a syringe.
• Approximately half of the aspirate should be placed in a blood collection tube containing either EDTA (purple top tube) or sodium heparin (green top tube). Use of an anticoagulant prevents clotting of the specimen and allows the laboratory to make smears at a later time. Anticoagulation also maximizes retrieval of bone marrows spicules.
• The other half of the aspirate should be allowed to clot in the syringe. After the marrow has clotted, the plunger should be removed from the syringe and the specimen remaining in the syringe poured into fixative. DO NOT eject the clot through the small end of the syringe using the plunger.
• Obtain bone marrow aspirate and biopsy and submit specimen for required studies as needed including tissue for routine microscopy, culture, chromosomal studies, and flow cytometry if indicated. The following specimens should be obtained as indicated:

• Bone core biopsy, Clotted aspirated material in a syringe for clot section preparation,

• Approximately 5 mL of aspirated material in a purple-top tube for aspirate smear preparation,

• Approximately 5 mL in a green-top (or purple-top or yellow-top) tube for flow cytometry (if ordered),

• Approximately 5 mL in a green-top tube for cytogenetic studies (if ordered).

• Culture material should be transmitted directly to a microbiology laboratory in an appropriate sterile container.
• Submit the specimens in containers labeled with the patient’s name, the requesting physician and the date the specimen was collected. If smears are prepared at the time of the procedure, slides must be labeled with the patient’s name. A properly completed specimen request form must accompany the specimens.
• After receipt in the laboratory and fixation for at least two hours, the core biopsy will be decalcified prior to further routine processing.

Tissue for Uric Acid Crystals

• Tissue for uric acid crystals should be submitted in 100% Alcohol or Blend Alcohol, NOT Formalin.
• Crystals are soluble in water and must NOT come in contact with water or any water-based fixative
• DO NOT put the specimen in Formalin
• Fix in Absolute (100%) Reagent-Absolute Isopropyl or Absolute Ethyl Alcohol only