Three-dimensional preparation and imaging reveal intrinsic microtubule properties.
We present an experimental investigation of microtubule dynamic instability in three dimensions, based on laser light-sheet fluorescence microscopy. We introduce three-dimensional (3D) preparation of Xenopus laevis egg extracts in Teflon-based cylinders and provide algorithms for 3D image processing. Our approach gives experimental access to the intrinsic dynamic properties of microtubules and to microtubule population statistics in single asters. We obtain evidence for a stochastic nature of microtubule pausing.
Keller
Philipp J
PJ
Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, D-69117 Heidelberg, Germany. keller@embl.de
Pampaloni
Francesco
F
Stelzer
Ernst H K
EH
D000818:Animals; D007091:Image Processing, Computer-Assisted / Q000379:methods; D021621:Imaging, Three-Dimensional / Q000295:instrumentation / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D008870:Microtubules / Q000502:physiology* / Q000648:ultrastructure*; D010063:Ovum / Q000648:ultrastructure; D014982:Xenopus laevis