Long-term two-photon fluorescence imaging of mammalian embryos without compromising viability

Jayne M. Squirrell*, David L. Wokosin, John G. White, Barry D. Bavister

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

488 Scopus citations

Abstract

A major challenge for fluorescence imaging of living mammalian cells is maintaining viability following prolonged exposure to excitation illumination. We have monitored the dynamics of mitochondrial distribution in hamster embryos at frequent intervals over 24 h using two-photon microscopy (1,047 nm) while maintaining blastocyst, and even fetal, developmental competence. In contrast, confocal imaging for only 8 h inhibits development, even without fluorophore excitation. Photo-induced production of H2O2 may account, in part, for this inhibition. Thus, two-photon microscopy, but not confocal microscopy, has permitted long-term fluorescence observations of the dynamics of three-dimensional cytoarchitecture in highly photosensitive specimens such as mammalian embryos.

Original languageEnglish (US)
Pages (from-to)763-767
Number of pages5
JournalNature biotechnology
Volume17
Issue number8
DOIs
StatePublished - Aug 1999

Keywords

  • Embryo
  • Hamster
  • Laser scanning confocal microscopy
  • Live cell fluorescence imaging
  • Mammal
  • Mitochondrial dynamics
  • Two-photon microscopy

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

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