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Confocal Laser Scanning Microscopy of Apoptosis in Organogenesis-stage Mouse Embryos by Robert M. Zucker; Sid Hunter; John M. Rogers is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Confocal Laser Scanning Microscopy of Apoptosis in Organogenesis-stage Mouse Embryos about?

Confocal laser scanning microscopy combined with a vital stain has been used to study apoptosis in organogenesis-stage mouse embryos. In order to achieve optical sectioning through embryos, it was necessary to use low power objectives and to prepare the sample appropriately. Mouse embryos were harvested on gestation day 8 or 9 and stained with the vital lysosomal dye, LysoTracker Red. Following incubation in the stain, embryos were fixed in 2% paraformaldehyde overnight, dehydrated in a graded methanol series, and cleared in benzyl alcohol/ benzyl benzoate. The resulting embryo is almost transparent and retains specific LysoTracker Red staining. The entire embryo can be optically sectioned and reconstructed in three dimensions to reveal areas of dye staining. To test this approach, the chemotherapeutic drug hydroxyurea was added to day 8 embryos in vitro to induce apoptosis. Our results demonstrated specific regions undergoing programmed cell death in normal development and increased apoptosis in embryos exposed to hydroxyurea. The observed patterns of LysoTracker Red staining correlate well with previous studies of cell death using other lysosomotropic dyes such as Nile blue sulfa

Who reads Confocal Laser Scanning Microscopy of Apoptosis in Organogenesis-stage Mouse Embryos?

Author

Robert M. Zucker; Sid Hunter; John M. Rogers

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Inc.; John Wiley & Sons Ltd.; Wiley (ISSN 0196-4763)

Published

1998

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)