CYTOPATHOLOGY

SPECIMEN COLLECTION PROCEDURES

GENERAL INFORMATION

1.         All cytological specimens should be considered infectious until fixed with a germicidal fixative.  Observe all mandated precautions when handling specimens for cytology.

2.         The following methods of specimen collection are intended to optimize cellular preservation for microscopic evaluation.  If there are any questions regarding the collection procedures, please contact the Cytology Department of the laboratory for further instructions.

REQUIREMENTS FOR ACCEPTABLE SPECIMENS

Slides and/or specimens are considered unacceptable if any one of the following exist:

1.         Any slide or specimen that is not labeled with the proper patient identification.  The patients last name and first initial is considered the minimum acceptable information.

2.         Any slide or specimen that is mislabeled or misidentified.  The name on the specimen and the Cytology Request form must match.

3.         Any slide or specimen that is improperly fixed or that is received in the improper container.

4.         Any slide or specimen that is not accompanied by a Cytology Request form.

5.         Any Cytology Request form lacking the following required minimum information:

(1)     Patient’s name

(2)     Patient’s age or date of birth

(3)     Name of the licensed physician, nurse practitioner or physician’s assistant who submitted the specimen.

(4)     Source of the specimen.

(5)     Date the specimen was collected.

(6)     Pertinent clinical history.

(A)  for Pap smears this should include date of last menstrual period and/or menstrual status.

CERVICAL AND VAGINAL CYTOLOGY (Pap Smears)

The ThinPrepÒ Pap Test and the traditional Pap smear are both evaluated by the laboratory. 

The ThinPrepÒ Pap Test is the preferred test due to its increased sensitivity in detecting LGSIL and more severe lesions.

Patient Preparation Considerations:

1.         The patient should not have douched or applied vaginal medication 24 hours prior to the examination.

2.         Preferably the patient should have the Pap smear taken two weeks after the first day of her last menstrual period and not when she is menstruating.

3.         The speculum should be inserted lubricated only with warm water. Lubricants should not be used.  Lubricants interfere with the slide preparation and result in unsatisfactory specimens.

4.         The collection of the Pap smear should be the first procedure in the gynecological examination.

Sample Collection:

1.         SPATULA

(1)      Insert the spatula into the cervix and rotate it 360° to perform the scraping.  Care should be taken to make sure the sample is taken from the transformation zone.

(2)      If the patient has had a hysterectomy, the spatula should be used to take the sample from the vaginal vault.

2.         ENDOCERVICAL BRUSH

(1)      Insert the endocervical brush into the endocervix with gentle pressure and rotate it only 180° to 360° to minimize bleeding.

3.         BROOM DEVICES

(1)      Insert the central bristles of the broom into the endocervical canal deep enough to allow the shorter bristles too fully contact the ectocervix.  Push gently, and rotate the broom in a clockwise direction five times.

Sample Preparation for Traditional Pap Smears:

1.         The sample should be spread uniformly across the surface of the slide.  It is important that the sample be applied to the slide before air-drying occurs.

2.         Immediately fix the smear by spraying with an aerosol fixative or coat the slide with the fixative provided in the Pap Pack.  It is important that the fixative is applied immediately to prevent any air-drying that will hinder the cytologic interpretation.

3.         Each slide should be properly labeled with the patient’s name.

Sample Preparation for ThinPrepÒ Pap Test:

1.         SPATULA

(1)      Rinse the spatula into the PreservCyt Solution vial by swirling the spatula vigorously in the vial 10 times.

2.         ENDOCERVIAL BRUSH

(1)      Rinse the brush in the PreservCyt Solution vial by rotating the brush in the solution 10 times while pushing against the vial’s bottom and wall.

3.         BROOM DEVICES

(1)      Rinse the broom into the PreservCyt Solution vial by pushing the broom into the bottom of the vial 10 times, forcing the bristles apart.  As a final step, swirl the broom vigorously to further release the material.

Tighten the cap so that the torque line on the cap passes the torque line on the vial.

Record the patient’s name on the vial.  The vial can now be sent to the lab for processing.

SPUTUM CYTOLOGY

1.         Have patient rinse mouth thoroughly with water.

2.         Obtain from a deep cough a thick mucoid specimen, expectorating it directly into a sputum fixative container.

(1)     The sputum fixative containers contain green colored Saccomanno fixative and are available from the laboratory.

(2)     Make sure the specimen is not saliva or nasopharyngeal secretions.

3.         Best results are achieved when three early morning sputa are collected on three consecutive days.  Send each specimen to the laboratory separately.

4.    If difficulty arises in obtaining a proper specimen, respiratory therapy can be used to induce an adequate sample.

URINARY TRACT CYTOLOGY

1.         Bladder washings should be collected in a sterile sealable container and sent to the lab for processing within six hours from the time of collection.  Do not add any fixative.

A.   If there will be more than a six hour delay before the specimen will be received by the lab add an equal amount of ThinPrepÒ PreservCyt Solution to the specimen..

2.         Urines should be collected in a sterile sealable container and sent to the lab for processing within six hours from the time of collection.  Do not add any fixative.

A.      If there will be more than a six hour delay before the specimen will be received by the lab add an equal amount of ThinPrepÒ PreservCyt Solution to the specimen.

B.       Do not use the morning’s first voided urine.

C.       If urine cytology times 3 is being collected, send each specimen separately to the laboratory after it has been collected.

D.   The specimen and request slip should specify whether the specimen was void or catheterized.

BODY FLUIDS, CSF & Miscellaneous CYST FLUIDS

1.         Collect fluid in a sterile sealable container.

2.    Send specimen immediately to the laboratory and refrigerate.

(1)     Do not add any fixative.

SKIN & MUCOUS MEMBRANE SCRAPINGS

1.         Scrape the site with a tongue blade or spatula.

(1)     Bullous lesions should be scraped from the base after removal of the roof.

2.         Rinse the sample into a PreservCyt Solution vial or a CytoLyt Solution vial by swirling the tongue blade or spatula vigorously in the vial 10 times.

(1)     PreservCyt Solution vials and CytoLyt Solution vials are available from the laboratory.

3.         Put the cap back on the vial.  Tighten the cap so that the torque line on the cap passes the torque line on the vial.

4.         Label the vial with proper patient identification and send it to the laboratory.

5.         If PreservCyt Solution or CytoLyt Solution vials are not available, transfer the specimen to a microscope slide and spray immediately with an aerosol coating fixative.

NIPPLE SECRETIONS

1.         Secretions from the nipple should be allowed to drop directly into a CytoLyt Solution vial.

(1)     CytoLyt Solutions vials are available from the laboratory.

2.         Put the cap back on the vial and tighten it so that the torque line on the cap passes the torque line on the vial.

3.         Label the vial with proper patient identification and send it to the laboratory.

4.         If CytoLyt solution vials are not available, smear the secretions on a microscope slide and spray immediately with an aerosol coating fixative.

ENDOSCOPIC BRUSHINGS & WASHINGS  (Bronchial & Gastrointestinal)

            Washings, Lavages & Aspirations:

1.         Physiologic saline or a balanced electrolyte solution, such as Plasma-LyteÒ or PolysolÒ, should be used for the washings.  Do not use water.

2.         Collect the specimen in a sterile sealable container.

3.         Send the specimen immediately to the laboratory and refrigerate.

(1)     Do not add any fixative.

Brushings:

1.         After the brushing has been taken, transfer the sample to a CytoLyt Solution vial.

(1)     Rinse the brush in the CytoLyt Solution vial by rotating the brush in the solution 10 times while pushing against the vial’s bottom and wall.

(2)     After rinsing the sample from the brush, cut the tip of the brush off and put it in the CytoLyt Solution vial.

2.         Do not reuse the brush.  The CytoLyt Solution contains methanol.

3.         Put the cap back on the vial.  Tighten the cap so that the torque line on the cap passes the torque line on the vial.

4.         Label the vial with proper patient identification and send to the laboratory.

FINE NEEDLE ASPIRATION BIOPSY

            General Considerations:

1.         A successful fine needle aspiration depends not only on performing the mechanical aspects of the procedure in a proper manner, but also on handling the specimen and preparing the smears in a manner that consistently produces quality samples for microscopic evaluation.  Contact the laboratory if there are any questions or if you wish to have a cytotechnologist present to help prepare the smears and handle the specimen.

2.         The type of preparation most appropriate for evaluating a given mass varies with the clinical situation, nature of the lesion and the preference of the pathologist.  Options include air-dried smears, fixed smears and preparation of the sample by ThinPrepÒ technique.  Bacteriologic studies may also be indicated.  Discussing the case with the pathologist prior to the procedure will optimize the preparation and the diagnostic results.    

Preparation of Smears:

1.         Smears are made from a small drop of semisolid aspirate placed on a glass slide. This is done by detaching the needle from the syringe and filling the syringe with air. Re-attach the needle and by advancing the plunger of the syringe express a small drop of aspirated material on the center of the slide.

2.         Make the smears by placing another slide on top of the first slide.  The slides are pulled apart with a sliding motion as the aspirate spreads from the weight of the slides.

3.         One smear should be air dried.  The other smear should be fixed immediately by spraying it with an aerosol fixative or by immersing the slide in 80% isopropanol.

4.         Make as many air-dried and fixed smears by this technique as the quantity of the aspirate permits.

Preparation by ThinPrepÒTechnique:

1.         Deposit and rinse the entire aspirate into a CytoLyt Solution vial.

(1)     CytoLyt Solution vials are available from the laboratory.

2.         Put the cap back on the vial and tighten it so the torque line on the cap passes the torque line on the vial.

3.         Label the vial with the proper patient identification and send it to the laboratory.