Regensburg 2007 – wissenschaftliches Programm

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BP: Fachverband Biologische Physik

BP 26: Poster Session II

BP 26.26: Poster

Donnerstag, 29. März 2007, 17:00–19:30, Poster B

Two-photon imaging and ablation of the mitotic spindle in S. pombe — •Nicola Maghelli and Iva Tolic-Norrelykke — MPI-CBG pfotenhauerstrasse 108 01307 DRESDEN

Multiphoton microscopy [1] has become a valuable tool for both in vitro and in vivo analysis of biological samples [2][3]. By focusing a near-infrared fs laser, it is possible to achieve photon densities high enough to exploit non-linear processes. The spatial volume in which such processes take places is around 0.5 µ m³; therefore confocal imaging is possible without the need of a pinhole. By increasing the power of the excitation laser, selective ablation inside living cells can be achieved [4]. We developed a custom-built two-photon microscope and applied it to study the effects of targeted nanosurgery inside the fission yeast S. pombe. Simultaneous ablation of the mitotic spindle and 3D imaging help understanding which forces are acting on the spindle during mitosis.
[1] Denk W, Strickler JH, Webb WW Two-photon laser scanning fluorescence microscopy. Science 1990 Apr 6;248(4951):73-6.

[2] Zipfel WR, Williams RM,WebbWWNonlinear magic: multiphoton microscopy in the biosciences. Nat Biotechnol. 2003 Nov;21(11):1369-77. Review.

[3] Squirrell JM, Wokosin DL, White JG, Bavister BD Long-term two-photon fluorescence imaging of mammalian embryos without compromising viability. Nat Biotechnol. 1999 Aug;17(8):763-7.

[4] Sacconi L, Tolic-Norrelykke IM, Antolini R, Pavone FS. Combined intracellular three-dimensional imaging and selective nanosurgery by a nonlinear microscope. J Biomed Opt. 2005 Jan-Feb;10(1):14002.