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Second Harmonic Imaging of Exocytosis at Fertilization

Department of Cell Biology and Center for Cell Analysis and Modeling, University of Connecticut Health Center, Farmington CT 06030-1507

Correspondence: Address reprint requests and inquiries to Leslie M. Loew, Tel.: 860-679-3568; E-mail: les{at}volt.uchc.edu.

Second harmonic generation offers an important alternative and complement to fluorescence for the imaging of cellular structure and function. Staining the eggs of the sea urchin, Lytechinus pictus, with the styryl dye di-8-ANEPPS, we have observed large changes in both second harmonic generation and two-photon fluorescence after fertilization, consistent with the dynamics of exocytosis of cortical granules. With nonlinear imaging on a scanning microscope, we are able to visualize the wave of exocytosis in real time.