Second Harmonic Imaging of Exocytosis at Fertilisation
Andrew C. Millard 1, Mark Terasaki 1 and Leslie M. Loew 1*
1 University of Connecticut Health Center
* To whom correspondence should be addressed. E-mail: les{at}volt.uchc.edu.
Submitted on March 29, 2005
Revised on April 17, 2005
Accepted on 19 April 2005
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Abstract |
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Second harmonic generation (SHG) 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 SHG and two-photon fluorescence following fertilisation, consistent with the dynamics of exocytosis of cortical granules. With non-linear imaging on a scanning microscope we are able to visualise the wave of exocytosis in real time.
Key Words:
cortical granules, exocytosis, microvilli, second harmonic imaging, styryl dye, two-photon fluorescence