Guidelines for Specimen Collection

We aim to deliver accurate and precise data to the medical community. However samples sometimes undergo changes between specimen collection and reporting that would result in inaccuracy of test results. Please refer to Table 1: Poor Specimen collection methods causing inaccurate test results to see the factors that cause inaccuracy.

One way of ensuring test results accuracy is to prevent anticoagulant contamination. To do this, the tubes should be filled in the order below:

Order of Draw:
i. Blood culture tubes (applying full aseptic technique)
ii. Red cap
iii. Blue cap
iv. Yellow cap
v. Green cap
vi. Purple cap
vii. Grey cap
viii. Royal Blue (Trace Element)

The caps of the tubes are colour coded to indicate the types of preservative or lack of preservative. Please refer to our BD Vacutainer Guide to view the type of preservative (or lack of it) in each tube.The tubes are provided free of charge by Gribbles Pathology.

Unless indicated, samples should be stored in room temperature (air-condition) and away from sunlight until pickup.

Department Guidelines  
Cytology Read
Histopathology Read
Microbiology Read

 



Cytology Guidelines

General Instructions on Specimen Fixation

  • Smears
    Rapid fixation of smears is necessary to preserve cytological detail. If smears are allowed to dry on the slides prior to fixation marked distortion of cells occurs.

    Smear preparations such as vaginal, cervical, endometrial aspirations and aspiration cytology specimens should be fixed immediately in a solution of 95% ethyl alcohol or a coating fixative such as Cytofix; other commercial spray fixatives may be used. A minimum of fifteen to twenty minutes fixation is required in the case of ethyl alcohol fixation (although prolonged fixation will not materially alter the cytology). In the case of spray fixatives, the smear should be allowed to dry for 10 minutes prior to placing into slide carries for despatch to the laboratory.
  • Fresh Specimens
    • •Specimens with high mucus content such as sputa and bronchial aspirates may be preserved for 12-24 hours if refrigerated.
    • • Specimens with high protein content such as pleural, peritoneal or pericardial fluid may be preserved for 24-48 hours with refrigeration.
    • • Specimens with low mucus or protein content such as urine or cerebrospinal fluid will endure 1-2 hours delay and so if delay is expected before delivery to the laboratory, fixative should be added to the specimen. The best in our experience is an equal volume of 50% ethyl alcohol.


Non- Gynaecological Specimens' Specific Instructions

  • Breast Cyst Aspiration – If aspirate is scanty, fluid is to be smeared one drop at a time on a clean slide and the slide is to be rapidly air dried. If aspirate is abundant, collect in a clean container and send fresh to the Cytology Department. Indicate the volume aspirated.
  • Breast Secretions (Nipple Discharge) – Drops of fluid from the nipple are smeared directly on clean glass slides. Submit 3 to 5 slides whenever possible. Half of the smears are to be fixed immediately with spray fixative or immersed in 95% alcohol for 20 minutes and another half of the smears should be left to air-dry without fixative. Clearly label all slides to indicate whether they are air dried or alcohol fixed.
  • Bronchial Brushings – Roll brush over clean, dry slide. Fix immediately with spray fixative or 95% ethyl alcohol. The brush(es) used to prepare bronchial brushing slides may be swished in a container of 70% ethyl alcohol to dislodge remaining specimen. Submit labeled slides and liquid together with the request form to the Lab.
  • Bronchial Washings – Collect in a clean container, label and send to the Lab.
  • Cerebrospinal Fluid – Collect 2 to 3 ml of CSF in a clean tube and send immediately to the Cytology Department (See also Part A. General Instructions on Specimen Fixation ii. Fresh Specimens above).
  • Colonic Washings – Collect in a clean container and send fresh to the Lab.
  • Effusions - Collect in a clean container and send fresh to the Lab.
  • Pelvic Washing - Collect in a clean container and send fresh to the Lab.
  • Fine Needle Aspiration – Fix 2 to 3 slides immediately (within a few seconds) using cytology spray fixative or immerse in 95% ethyl alcohol for 20 minutes and rapidly air dry 2–3 slides without fixative. If fluid is obtained with a needle pass, it should be expressed into a clean container. Clearly label container and all slides to indicate which is air dried and which is alcohol fixed. Submit to the Lab using one request form.


Gynaecological specimens

Cervical smears (Conventional method)
Routine smear which combines ectocervical and endocervical canal scrappings are most satisfactory.
Spatula for ectocervical samples and a cytobrush for the endocervix are recommended.

SurePath Pap Test (Liquid – based)
The SurePath Pap Test is being used as a replacement for the Pap smear. The SurePath Pap Test overcomes the limitations of the conventional method. By improving the way sample slide is prepared, the SurePath Pap Test actually improves the quality of the test.

The BD SurePath™ liquid-based Pap test is an FDA approved thin-layer cell preparation process intended for use in the screening and detection of cervical cancer, pre-cancerous lesions, atypical cells and all other cytologic categories as defined by The Bethesda System for Reporting Cervical/Vaginal Cytologic Diagnoses.

  • With SurePath liquid-based Pap test increases detection by 64.4% in HSIL+ lesions comparison to the conventional Pap smear.
  • Significantly reduces Unsatisfactory cases for potentially fewer patient call-backs.
  • Allows physician to perform Pap tests in a wide variety of clinical presentations (e.g. blood, mucous and inflammation) with maximum confidence in the result due to the proprietary SurePath Cell Enrichment process used in the lab.


SurePath Pap test has a unique, easy to use collection process that standardizes the collection process and ensures 100% of the collected sample is sent to the laboratory for processing. No diagnostic cells are discarded in the clinic, as is true with “rinse and swish” systems, which on average can result in the loss of on average, 37% of the collected sample during cell transfer.

Patient Preparation for Cervical smear
Proper patient preparation encompasses the following:
Ideal sampling date is two weeks after the first day of the LMP.
Discourage sampling during normal menses.
Avoid use of vaginal medication, vaginal contraceptives, or douches for 48 hours prior to examination.

Guidelines for Referral of Specimen
The specimen must be received in an appropriately labeled container.
The specimen must be accompanied with a General Request form, with appropriate and relevant clinical data.
Details of treatment (e.g. HRT, DXT) and procedures (e.g. LLETZ, Cone biopsy) must be included in the clinical data.


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Histopathology Guidelines
A test request form must accompany all specimens. When more than one specimen is sent from the same patient at the same operation, please use only one form. Specimens from different anatomical sites should be sent in separate containers and must be itemised in the same request form. The containers of such samples must similarly be itemised and labeled with patient’s identification so that they can be cross–referenced to the patient and the anatomic site of origin of the sample.

The request form must include all that is required as in the General Guidelines to filling the General Request Form. In addition to that, please ensure the details below are also provided:

  •   Examination required e.g. cytology, histology etc.
  •   Previous biopsy number / diagnosis, if any.
  •   Name and signature of physician/ surgeon in charge of case.

 Special Instructions on Specimen Collection & Handling

Routine specimens should be sent in containers with 10% buffered formalin unless otherwise stated.

Appropriate containers are available from the laboratory on request. To prevent leakage it is advised to double wrap the specimen container.

Multiple small specimens, such as gastrointestinal biopsies, should be mounted on a piece of filter paper and properly labeled.

Large specimens such as colon, should be opened, the contents cleaned out and specimen should be completely immersed in formalin. Containers must be tightly secured.

Do not crush specimens with forceps, hemostats or other instruments. Avoid using cautery.

Do not force a large specimen into a small container. Large specimens must be completely surrounded by formalin for proper fixation.

Please tick the “URGENT” box on the General Request form; otherwise, the specimen will be processed as per normal.

For specimens where orientation is important, mark or tag the specimen e.g. Axillary tail of mastectomy specimens, orientation of surgical margin.


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Microbiology Guidelines
Please refer to Table 2: Guidelines for Microbiology Specimen Collection

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