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MBC Video for Molecular Biology of the Cell: Volume 0, Issue 2003 -- E02-04-0236

QuickTime Videos

• Figure1_movie (5.77 MB)

  Labeled EB1 binds polymerizing ends of astral and spindle microtubules, and a region within the kinetochore attachment site. Alexa594-labeled EB1 microinjected into a mitotic PtK1 cell and imaged by spinning disk confocal microscopy in a single focal plane every ten seconds. Polymerizing astral and spindle microtubules as well as spindle poles show EB1 localization in the characteristic comet pattern. Both spindle poles are visible at the start of the time-lapse; the lower pole subsequently rotates out of the focal plane. The dark shadows in the central spindle are areas where EB1 is excluded by chromosomes. Time acceleration, 40x.

• Figure5A_Metaphase_EB1.mov (4.77 MB)

  EB1 binds polymerizing kinetochore microtubule ends on bi-oriented chromosomes. Alexa594-labeled EB1 microinjected into a metaphase PtK1 cell; paired fluorescence and phase contrast images acquired by spinning disk confocal microscopy in a single focal plane every seven seconds. EB1 fluorescence (green) is superimposed on a phase contrast image to show the chromosomes (red). EB1 is seen on anti-poleward moving kinetochores, corresponding to kinetochore microtubule polymerization. The lower spindle pole is visible, while the upper pole is out of the plane of focus. Arrows show some examples of EB1 at anti-poleward moving kinetochores. Time acceleration, 35x.

• Figure5D_Monastrol_treated_EB1.mov (5.44 MB)

  EB1 binds polymerizing kinetochore microtubule ends on mono-oriented chromosomes. Alexa594-labeled EB1 microinjected into a PtK1 cell pretreated with Monastrol. Paired fluorescence and phase contrast images were acquired by spinning disk confocal microscopy in a single focal plane every seven seconds in the presence of Monastrol. EB1 fluorescence (green) is superimposed phase contrast (red, to show chromosomes). Monastrol treatment induces monoastral spindles encircled by oscillating chromosomes. The spindle poles are labeled in the first frame. EB1 is seen on anti-poleward moving kinetochores, corresponding to kinetochore microtubule polymerization. Rarely, both kinetochores of a pair show EB1 fluorescence. Arrows show selected examples of EB1 targeting to kinetochores. Time acceleration, 35x.

• Figure5E_Anaphase_EB1.mov (2.81 MB)

  EB1 binds polymerizing kinetochore microtubule ends on segregating sister chromosomes during anaphase. Alexa594-labeled EB1 microinjected into a metaphase PtK1 cell. Paired fluorescence and phase contrast images were acquired through anaphase chromosome segregation by spinning disk confocal microscopy in a single focal plane every seven seconds. EB1 fluorescence (green) is superimposed on phase contrast (red, to show chromosomes). The time-lapse begins during metaphase and continues through the end of anaphase. Kinetochores move poleward during the majority of anaphase, but during rare anti-poleward oscillations, corresponding to microtubule polymerization, EB1 is seen at kinetochores. Arrows show two examples of this, for a chromosome associated with the lower spindle pole. Time acceleration, 35x.

Prepared by: the MBC Journals Production Manager

This Article Abstract Full Text Services Alert me to new issues of the journal