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Page Title: Tritiated Thymidine BeLPT: Bronchoalveolar Lavage (BAL) Specimen
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DOE-SPEC-1142-2001
7. TRITIATED THYMIDINE BeLPT: BRONCHOALVEOLAR LAVAGE (BAL)
SPECIMEN
7.1.
SAMPLE COLLECTION. For the BAL-LPT, the specimen is a saline lavage
obtained during bronchoscopy. It is recommended that samples be taken from the
wedged position in lingula or right middle lobe. The lavage is preformed before the
biopsy to avoid contamination with blood. A minimum of 480 ml normal saline is
instilled in two 240 ml aliquots. Gentle aspiration occurs after each aliquot is
instilled. Fluid recovery usually varies from 50-70%. The specimen must be
transported to the laboratory immediately. Temperature should be kept between 15 to
25C using insulation if necessary.
Specimens that cannot be set up for proliferation testing that day must be suspended
in RPMI-1640 with antibiotics and processed within 36 hours (no more than one
night should pass before the cells are placed into culture for proliferation assays). The
test requires a minimum of 2.5 x 106 cells and 20 x 106 are recommended.
7.2.
SAMPLE PREPARATION. BAL preparations from normal individuals usually
consist of over 90% alveolar macrophages. However, in beryllium disease higher
percentages of lymphocytes are typically observed. BAL fluid is centrifuged, the cells
re-suspended in culture medium, counted, and exposed to beryllium for 3 and 5 days.
The presence of immuno-competent T lymphocytes in the preparation is usually
ascertained by testing with Candida albicans, a common recall antigen, although
comparable universal antigens/mitogens may be employed to the same end. Tritiated
thymidine is included for the last 6-18 hrs. of culture and the assay is terminated by
harvesting cells onto filter paper and determining the radioactive content in a
scintillation or gas ionization counter.
7.3.
SAMPLE FORM
7.3.1. PATIENT NAME: BIRTH DATE:
7.3.2. HOSPITAL#
7.3.3. DATE AND TIME OF TEST: SW:
7.3.4. SPECIMEN #
7.3.5. LAVAGE VOLUME- ml IN:
7.3.6. ml OUT: %Return:
7.3.7. CELL YIELD: Total number of cells recovered/ml lavage fluid out
6


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