SEPARATION OF LYMNAEA-STAGNALIS HEMOCYTES BY DENSITY GRADIENT CENTRIFUGATION

A chelating anti-clumping (alpha-C) buffer allowed blood cells (hemocy tes) of a gastropod, Lymnaea stagnalis to be separated by discontinuou s Percoll density gradient centrifugation. The hemocytes of L. stagnal is were separated into five fractions, having a density lower than 10, 20, 30, 40, and 50% Percoll, respectively. Trypan blue exclusion assa ys showed viability of separated hemocytes to be between 81 and 89%. C ytospin preparations of these hemocytes were examined. Small cells wer e mainly observed at high densities; at lower densities medium and lar ge hemocytes were also present. No absolute separation was achieved. S ome density fractions were enriched for hemocytes with regard to the d istributions of two endogenous lysosomal enzymes (alpha-naphthyl aceta te esterase and acid phosphatase).