PMID: 9551178Jan 1, 1997Paper

High-resolution imaging at the cellular and subcellular levels in flattened whole mounts of early zebrafish embryos

Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire
T D YagerD Brooker

Abstract

We describe a rapid and sensitive method for high-resolution imaging at the cellular and subcellular levels in the whole-mount zebrafish embryo. The procedure involves fixing and staining the embryo, followed by deyolking and flattening it under a cover slip, to produce a planar mount that is 20 to 100 microns thick. Such a flattened whole mount allows imaging with a spatial resolution of approximately 500 nm in the x-y plane and does not require the use of embedding, sectioning, confocal microscopy, or computational deblurring procedures. We can resolve all individual nuclei and chromosome sets in the embryo, up to the late gastrula stage (10,000 cell stage). In addition, older embryos (through the segmentation stage) can also be examined, with the preservation of significant morphological detail. Because of its ability to resolve subcellular detail, the flattened whole-mount method can provide significant biological information beyond what can be obtained from conventional (three-dimensional) whole mounts. We have used the flattened whole-mount method to study subcellular events related to progression through the cell cycle or to apoptosis, in cells of the early zebrafish embryo. A specific DNA-binding dye (Hoechst 33258) or ...Continue Reading

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Citations

Jun 19, 2001·Proceedings of the National Academy of Sciences of the United States of America·P A Padilla, M B Roth
Jun 25, 2005·Developmental Dynamics : an Official Publication of the American Association of Anatomists·R Craig AlbertsonPamela C Yelick
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