THE EFFECT OF LABELING INTENSITY, ESTIMATED BY REAL‐TIME CONFOCAL LASER SCANNING MICROSCOPY, ON FLOW CYTOMETRIC APPEARANCE AND IDENTIFICATION OF IMMUNOCHEMICALLY LABELED MARINE DINOFLAGELLATES1

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Abstract
Two different fluorescein isothiocyanate (FITC) conjugates were used to analyze the effect of labeling intensity on the flow cytometric appearance of marine dinoflagellates labeled with antibodies that specifically recognized the outer cell wall. Location of the labeling was revealed by epifluorescence and real‐time confocal laser scanning microscopy using an anti‐rabbit IgG/FITC‐conjugated secondary antiserum. Flow cytometric measurements showed that cells ofProrocentrumspecies labeled this way could not always be distinguished from unlabeled cells. The labeling intensity increased several times when a biotinylated anti‐rabbit IgG secondary antiserum was used in combination with a streptavidin/FITC conjugate. Flow cytometry indicated that the labeling intensity had increased 50%, which resulted in an improved separation of clusters of labeled and unlabeled cells.