Flow cytometric analysis of lectin-stained haemocytes of the honeybee (Apis mellifera)Dirk C. de Graafa, Rebecca Dauweb, Karl Walravensa and Frans J. Jacobsb
a Veterinary and Agrochemical Research Centre, Groeselenberg 99, 1180 Brussels, Belgium
b University of Ghent, Laboratory of Zoophysiology, Krijgslaan 281 S33, 9000 Ghent, Belgium
(Received 18 July 2001; revised 18 February 2002; accepted 18 April 2002)
The study evaluated the technical feasibility of distinguishing the different honeybee blood cells by flow cytometric analysis with and without staining by 3 lectins: soybean agglutinin (SBA), concanavalin A (ConA) and wheat germ agglutinin (WGA). Flow cytometric analysis of unstained cells provided a forward scatter-side scatter dotplot with no distinct haemocyte population. Examination of blood samples stained with FITC-labelled ConA or WGA revealed a notable population of fluorescently marked cells on FL1-histograms. Microscopic analysis, run parallel, demonstrated low fluorescence of the granular cells, strong fluorescence of the plasmatocytes, with P1's stained all over their surfaces and P2's with a rather dotted appearence. Prohaemocytes were not stained at all. SBA-FITC did not stain honeybee haemocytes as demonstrated by both the flow cytometric and the microscopic examinations.
Key words: honeybee / haemocytes / flow cytometry / lectin staining
Correspondence and reprints: Dirk C. de Graaf
© INRA, EDP Sciences, DIB, AGIB 2002