Localization of the major allergen Bet v 1 in birch pollen by confocal laser scanning microscopy

Confocal laser scanning microscopy (CLSM) was used to localize the major allergen, Bet v I in birch pollen. Pollen grains of Betula pendula were collected from catkins that had begun to release their pollen, freeze dried, and stored at 4 degrees C. The pollen material was cryosectioned and stained with indirect immunofluorescence using the monoclonal antibody Bv 10. The observed localization of Bet v I was dependent on the treatment of the grains. When they had been prefixed with gaseous formaldehyde followed by acetone Bet v I was found in the cytoplasm. When the prefixation was omitted, a minor portion of Bet v I also appeared in the exine in the aperture regions. The allergen was not observed in the inline or the Zwischenkorper. Our suggestion is that the normal route for excretion of Bet v I is via the apertures on contact between pollen and the stigmatic surface of the pistil.