Separation of Lymnaea stagnalis hemocytes by density gradient centrifugation.
A chelating anti-clumping (alpha-C) buffer allowed blood cells (hemocytes) of a gastropod, Lymnaea stagnalis to be separated by discontinuous Percoll density gradient centrifugation. The hemocytes of L. stagnalis were separated into five fractions, having a density lower than 10, 20, 30, 40, and 50% Percoll, respectively. Trypan blue exclusion assays showed viability of separated hemocytes to be between 81 and 89%. Cytospin preparations of these hemocytes were examined. Small cells were mainly observed at high densities; at lower densities medium and large hemocytes were also present. No absolute separation was achieved. Some density fractions were enriched for hemocytes with regard to the distributions of two endogenous lysosomal enzymes (alpha-naphthyl acetate esterase and acid phosphatase).
Adema
C M
CM
Department of Medical Microbiology and Parasitology, Faculty of Medicine, Vrije Universiteit, Amsterdam, The Netherlands.
Mohandas
A
A
van der Knaap
W P
WP
Sminia
T
T
D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D002469:Cell Separation / Q000379:methods*; D048429:Cell Size; D002470:Cell Survival; D002499:Centrifugation, Density Gradient / Q000379:methods*; D006434:Hemocytes / Q000166:cytology* / Q000201:enzymology; D008195:Lymnaea / Q000166:cytology* / Q000201:enzymology; D008247:Lysosomes / Q000201:enzymology; D009283:Naphthol AS D Esterase / Q000378:metabolism; D011205:Povidone; D012822:Silicon Dioxide
C016039:Percoll; D012822:Silicon Dioxide; D009283:Naphthol AS D Esterase; D000135:Acid Phosphatase; D011205:Povidone