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Confocal Laser Scanning Microscopy of Whole Mouse and Human Embryos: Apoptosis and Morphology.

Published online by Cambridge University Press:  02 July 2020

Zucker Robert M.
Affiliation:
Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, Research Triangle Park, NC. 27711.
Sulik Kathy
Affiliation:
Department of Cell Biology and Anatomy. The University of North Carolina, Chapel HillNC
Owen T Price
Affiliation:
Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, Research Triangle Park, NC. 27711.
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Extract

Two novel sample preparation procedures have been developed to visualize morphological structures in embryonic and fetal tissues. In the first, optimization of the sample preparation allows for the visualization of embryos with a thickness in excess of 500 microns. The morphology of the internal structures in these embryos is observed by optical sectioning. The general sample preparation procedures included paraformaldehyde fixation, methanol dehydration and clearing with either benzyl alcohol/benzyl benzoate or methyl salicylate. The dyes (i.e. LysoTracker Red, YoPro) are incorporated into living or fixed embryos to indicate a specific biological function or embryo morphology. Using these stained embryos, the detection of apoptosis and its subsequent quantification were made in addition to the observation of morphology.

In the second technique, the confocal microscope has been adapted to produce images similar to those generated by a scanning electron microscope. For this application, the embryos are coated with a dilute solution of acridine orange resulting in opaque surface fluorescence.

Type
Applications of Imaging Techniques to the Study of Embryological Development
Copyright
Copyright © Microscopy Society of America

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References

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