Preferred volume: In a sterile transport container such as a 50mL Falcon tube (352070), place about 10-20mL of Bronchoalveolar Lavage (BAL) fluid.
Exact volume required is dependant on lymphocyte count.
Transport to lab as soon as possible after collection. If transport will be delayed, keep specimen on ice to prevent bacterial overgrowth.
Follow institution's bronchoscopy protocol.
Minimum volume required is dependent on cell count. Any volume >3mL will be accepted and tested for lymphocyte count. If sufficient lymphocytes are present (>10% on differential and => 1 million lymphocytes total) phenotyping will be performed.
Frozen or visibly hemolyzed samples. Samples >24 hours old will have a viability test performed. If <50% of cells are viable, phenotyping will not be performed.
Ambient if transport time is under 1 hour. Place on ice for transport times between 1 and 4 hours. For transport times >4 hours, see Special Handling Instructions under Notes.
1 hour ambient, 4 hours refrigerated, 24 hours refrigerated in supplemented cell culture media.
1. Lymphocyte phenotyping will not be performed on samples with <10% lymphocytes and/or 1 million total lymphocytes.
2. Cell count and differential (LAVCT) must also be ordered.
3. Samples have spedific handling instructions and shipped specimens require special transport media.
4. Volumes of fluid instilled and recovered must accompany the sample to calculate absolute cell counts.
5. Phenotyping includes: CD3, CD4, CD8, CD4:CD8 ratio and CD19.
Special Handling Instructions:
1. Record original volumes (instilled and recovered). Must be included with sample.
2. Centrifuge sample at 500 x g for 5 minutes.
3. Discard supernatant and gently resuspend cell pellet with the same volume of supplemented cell culture media such as RPMI + 1% L-glutamine, 1% Penicillin/Streptomycin and 10% Fetal Bovine Serum (FBS).
4. Ship sample on cold pack via Fed Ex Priority Overnight Monday through Thursday only.
Immunology Lab - Phenotyping Assays
BAL or Lavage Phenotyping
86360, 86355, 86359