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UV microbeam irradiations of the mitotic spindle I. The UV-microbeam apparatus

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Summary

We describe the assembly of a UV microbeam microscope based on a Zeiss IM35 inverted microscope. The important UV transmitting elements are standard UV epifluorescence attachments available from Zeiss; the main modification involves fitting an adjustable slit in place of the field diaphragm. We describe how to align and focus the UV source for optimal irradiations. Our current version of this machine is also fitted with a monochromator and using monochromatic UV light, we can reproduceably create Areas of Reduced Birefringence in spindle fibres with ca. 2–3 s irradiations, while continually observing the fibres. The microscope is stable and easy to set up, allowing many consecutive experiments to be done, including multiple irradiations on the one cell. In conjunction with video image processing techniques, the cells can be observed continuously using polarising, Nomarski or other optical systems. Some preliminary observations demonstrating the versatility of the machine are described.

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Abbreviations

ARB:

areas of reduced birefringence

MT:

microtubules

UV:

ultraviolet

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Author notes

  1. J. D. Pickett-Heaps

    Present address: School of Botany, University of Melbourne, 3052, Parkville, Victoria, Australia

  2. O. G. Stonington

    Present address: The Colorado Rocky Mountain School, 1493 County Rd. 106, 81623, Carbondale, CO, USA

  3. Judith A. Snyder

    Present address: Department of Biological Sciences, University of Denver, 80208, Denver, CO, USA

Authors and Affiliations

  1. Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder, Colorado

    O. G. Stonington, T. P. Spurck, Judith A. Snyder & J. D. Pickett-Heaps

Authors
  1. O. G. Stonington
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  2. T. P. Spurck
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  3. Judith A. Snyder
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  4. J. D. Pickett-Heaps
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Stonington, O.G., Spurck, T.P., Snyder, J.A. et al. UV microbeam irradiations of the mitotic spindle I. The UV-microbeam apparatus. Protoplasma 153, 62–70 (1989). https://doi.org/10.1007/BF01322466

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  • DOI: https://doi.org/10.1007/BF01322466

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