Controlled damage in thick specimens by multiphoton excitation

James Galbraith, Mark Terasaki

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Controlled damage by light energy has been a valuable tool in studies of cell function. Here, we show that the Ti:Sapphire laser in a multiphoton microscope can be used to cause localized damage within unlabeled cells or tissues at greater depths than previously possible. We show that the damage is due to a multiphoton process and made wounds as small as 1 μm in diameter 20 μm from the surface. A characteristic fluorescent scar allows monitoring of the damage and identifies the wound site in later observations. We were able to lesion a single axon within a bundle of nerves, locally interrupt organelle transport within one axon, cut dendrites in a zebrafish embryo, ablate a mitotic pole in a sea urchin egg, and wound the plasma membrane and nuclear envelope in starfish oocytes. The starfish nucleus collapsed ∼1 h after wounding, indicating that loss of compartmentation barrier makes the structure unstable; surprisingly, the oocyte still completed meiotic divisions when exposed to maturation hormone, indicating that the compartmentalization and translocation of cdk1 and its regulators is not required for this process. Multiphoton excitation provides a new means for producing controlled damage deep within tissues or living organisms.

Original languageEnglish (US)
Pages (from-to)1808-1817
Number of pages10
JournalMolecular Biology of the Cell
Volume14
Issue number5
DOIs
StatePublished - May 1 2003
Externally publishedYes

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Starfish
Oocytes
Axons
Wounds and Injuries
Sea Urchins
Aluminum Oxide
Nuclear Envelope
Zebrafish
Dendrites
Organelles
Cicatrix
Ovum
Lasers
Embryonic Structures
Cell Membrane
Hormones
Light

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

Controlled damage in thick specimens by multiphoton excitation. / Galbraith, James; Terasaki, Mark.

In: Molecular Biology of the Cell, Vol. 14, No. 5, 01.05.2003, p. 1808-1817.

Research output: Contribution to journalArticle

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