Controlled cultivation of Alexandrium minutum and [P-33] orthophosphate cell labeling towards surface adhesion tests

We describe a protocol for preparing cultures of the PSP (paralytic shellfi sh poisoning) dinoflagellate Alexandrium minutum, towards subsequent studie s of cell adhesion onto artificial substrates. First, phenotypic uniformity of the strain used and reproducibility of the standard growth profile are obtained by optimising key parameters. Batches of A. minutum at mid-exponen tial proliferation phase are radiolabeled with P-33-containing medium in or der to later quantify cell adhesion. ii mortality corrective index is appli ed to the latter, using the vital fluorochrome acridine orange, i.e. dead c ells show no nuclear incorporation under epifluorescence microscopy. (C) 19 99 Elsevier Science B.V. All rights reserved.