Bronchoalveolar lavage fluid processing. Effect of membrane filtration preparation on neutrophil recovery

Two common methods for the preparation of bronchoalveolar lavage (BAL) fluid for cytologic examination, cytocentrifugation and membrane filtration, have been found to yield different results in the quantitation of lymphocytes. To compare these two methods for the quantitation of neutrophils, the differential counts from 640 consecutive clinical specimens were analyzed retrospectively. The percentage of neutrophils resulting from the preparation of the BAL fluids by the two methods were highly correlated (r2 = .72). However, cytocentrifugation yielded consistently higher neutrophil percentages than did membrane filtration (means for all samples: 18.5 +/- 1.0% vs. 14.7 +/- 0.9%; P less than .001). To investigate the source of the variation in neutrophil quantitation by the two methods, two series of mixing experiments were performed in which neutrophil-rich cell suspensions were added to BAL fluids. Determination of the cellular differentials before and after mixing the cell suspensions demonstrated that membrane filtration preparation tends to lose neutrophils while cytocentrifugation accurately recovers neutrophils. Thus, accurate quantitation of the two cells recovered by BAL may require use of both cytocentrifugation and membrane filtration.