Human Reproduction, Vol 12, 1250-1252, Copyright © 1997 by Oxford University Press
D Keefe, P Tran, C Pellegrini and R Oldenbourg
The zona pellucida (zona) is a glycoprotein coat that envelopes the oocyte
and embryo, binds sperm during fertilization and facilitates transfer of
the embryo through the Fallopian tube. Before implantation can occur, the
blastocyst must hatch from the zona. Several lines of evidence suggest that
the zona is multilaminar. We hypothesized that the multilaminar structure
of the zona filaments could be imaged non- destructively with the polarized
light microscope. A recent modification of the polarized light microscope
(pol-scope), which combines innovations in polarization optics with novel
image processing software, allows measurement of birefringence at all
points of the image. Hamster metaphase II oocytes were placed on glass
coverslips which replaced the bottom of culture dishes, imaged under
differential interference contrast (DIC) and pol-scope optics, then
digitized and processed to measure birefringence magnitude and orientation.
The pol- scope revealed the zona to be divided into outer and inner layers
separated by a zone of low retardance. This finding is consistent with
filaments in the outer layer oriented tangentially and in the inner layer
oriented radially. The multilaminar structure of the mammalian zona
suggested by differential lectin binding and by scanning electron
microscopy could be imaged non-destructively with the pol-scope. Because
the pol-scope provides a non-destructive method to identify macro-molecular
organization of the zona, it may prove useful in developmental studies of
hatching and to direct resection of the zona.
ARTICLES
Polarized light microscopy and digital image processing identify a multilaminar structure of the hamster zona pellucida
Department of Obstetrics and Gynecology, Women and Infants Hospital, Brown University School of Medicine, Providence, RI 02905, USA.
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