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Analysis of Microtubule Dynamics by Polarized Light

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Microtubule Protocols

Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 137))

Abstract

In this article, the author describes the needed instrumentation and the methods to be followed for the observation and measurement of the birefringence of single and bundled microtubules and of their ordered arrays using a polarizing microscope. As instruments, the traditional polarizing microscope and the recently developed LC-PolScope are discussed. As methods, we describe qualitative and quantitative observations, including notes on specimen preparations, which optimize the sensitivity and accuracy of measuring specimen retardance.

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References

  1. Inoué, S. (1952) The effect of cholchicine on the microscopic and submicroscopic structure of the mitotic spindle. Exp. Cell Res. Suppl. 2, 305–318.

    Google Scholar 

  2. Inoué, S. (1964) Organization and function of the mitotic spindle, in Primitive Motile Systems in Cell Biology, (Allen, R. H. and Kamiya, N., eds.), Academic Press, New York pp. 549–598.

    Google Scholar 

  3. Inoué, S. and Sato, H. (1967) Cell motility by labile association of molecules: the nature of mitotic spindle fibers and their role in chromosome movement. J. Gen. Physiol. 50, 259–292.

    Article  PubMed  Google Scholar 

  4. Salmon, E. D. (1975) Pressure-induced depolymerization of spindle microtubules. I. Changes in birefringence and spindle length. J. Cell Biol. 65, 603–614.

    Article  CAS  PubMed  Google Scholar 

  5. Sato, H., Ellis, G. W., and Inoué, S. (1975) Microtubular origin of mitotic spindle form birefringence Demonstration of the applicability of Wiener’s equation. J. Cell Biol. 67, 501–517.

    Article  CAS  PubMed  Google Scholar 

  6. Inoué, S. and Salmon, E. D. (1995) Force generation by microtubule assembly/disassembly in mitosis and related movements. Mol. Biol. Cell 6, 1619–1640.

    PubMed  Google Scholar 

  7. Inoué, S. and Hyde, W. L. (1957) Studies on depolarization of light at microscope lens surfaces II. The simultaneous realization of high resolution and high sensitivity with the polarizing microscope. J. Biophys. Biochem. Cytol. 3, 831–838.

    Article  PubMed  Google Scholar 

  8. Inoué, S. and Kubota, H. (1958) Diffraction anomaly in polarizing microscopes. Nature 182, 1725–1726.

    Article  PubMed  Google Scholar 

  9. Oldenbourg, R. and Mei, G. (1995) New polarized light microscope with precision universal compensator. J. Microsc. 180, 140–147.

    CAS  PubMed  Google Scholar 

  10. Shribak, M. and Oldenbourg, R. (2003) Techniques for fast and sensitive measurements of two-dimensional birefringence distributions. Appl. Opt. 42, 3009–3017.

    Article  PubMed  Google Scholar 

  11. Oldenbourg, R. (2005) Polarization microscopy with the LC-PolScope, in Live Cell Imaging: A Laboratory Manual, (Goldman, R. D. and Spector, D. L., eds.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 205–237.

    Google Scholar 

  12. LaFountain, J. R., Jr. and Oldenbourg, R. (2004) Maloriented bivalents have metaphase positions at the spindle equator with more kinetochore microtubules to one pole than to the other. Mol. Biol. Cell 15, 5346–5355.

    Article  CAS  PubMed  Google Scholar 

  13. Liu, L., Oldenbourg, R., Trimarchi, J. R., and Keefe, D. L. (2000) A reliable, noninvasive technique for spindle imaging and enucleation of mammalian oocytes. Nat. Biotechnol. 18, 223–225.

    Article  CAS  PubMed  Google Scholar 

  14. Wang, W. H. and Keefe, D. L. (2002) Prediction of chromosome misalignment among in vitro matured human oocytes by spindle imaging with the PolScope. Fertil. Steril. 78, 1077–1081.

    Article  PubMed  Google Scholar 

  15. Sammak, P. J. and Borisy, G. G. (1988) Direct observation of microtubule dynamics in living cells. Nature 332, 724–726.

    Article  CAS  PubMed  Google Scholar 

  16. Horio, T. and Hotani, H. (1986) Visualization of the dynamic instability of individual microtubules by dark-field microscopy. Nature 321, 605–607.

    Article  CAS  PubMed  Google Scholar 

  17. Walker, R. A., O’Brien, E. T., Pryer, N. K., et al. (1988) Dynamic instability of individual microtubules analyzed by video light microscopy: rate constants and transition frequencies. J. Cell Biol. 107, 1437–1448.

    Article  CAS  PubMed  Google Scholar 

  18. Oldenbourg, R., Salmon, E. D., and Tran, P. T. (1998) Birefringence of single and bundled microtubules. Biophys. J. 74, 645–654.

    Article  CAS  PubMed  Google Scholar 

  19. Waterman-Storer, C. M. (1998) Microtubules and microscopes: how the development of light microscopic imaging technologies has contributed to discoveries about microtubule dynamics in living cells. Mol. Biol. Cell 9, 3263–3271.

    CAS  PubMed  Google Scholar 

  20. Cassimeris, L., Pryer, N. K., and Salmon, E. D. (1988) Real-time observations of microtubule dynamic instability in living cells. J. Cell Biol. 107, 2223–2231.

    Article  CAS  PubMed  Google Scholar 

  21. Inoué, S. (2002) Polarization microscopy, in Current Protocols in Cell Biology, (Bonifacino, J. S., Dasso, M., Harford, J. B., Lippincott-Schwartz, J., and Yamada, K. M., eds.), John Wiley & Sons, New York, pp. 4.9.1–4.9.27.

    Google Scholar 

  22. Inoué, S. (1986) Video Microscopy. Plenum Press, New York.

    Google Scholar 

  23. Oldenbourg, R. (1999) Polarized light microscopy of spindles. Methods Cell Biol. 61, 175–208.

    Article  CAS  PubMed  Google Scholar 

  24. Shribak, M., Inoué, S., and Oldenbourg, R. (2002) Polarization aberrations caused by differential transmission and phase shift in high NA lenses: theory, measurement and rectification. Opt. Eng. 41, 943–954.

    Article  Google Scholar 

  25. Oldenbourg, R. (1996) A new view on polarization microscopy. Nature 381, 811–812.

    Article  CAS  PubMed  Google Scholar 

  26. Inoué, S. and Dan, K. (1951) Birefringence of the dividing cell. J. Morphol. 89, 423–456.

    Article  Google Scholar 

  27. Inoué, S. and Sato, H. (1966) Deoxyribonucleic acid arrangement in living sperm, in Molecular Architecture in Cell Physiology, (Hayashi, T. and Szent-Gyorgyi, A. G., eds.), Prentice Hall, Englewood Cliffs, NJ, pp. 209–248.

    Google Scholar 

  28. Inoué, S. and Spring, K. R. (1997) Video Microscopy. Plenum Press, New York.

    Google Scholar 

  29. Ihoué, S. and Spring, K. R. (1997) Video Microscopy. Plenum Press, New York.

    Google Scholar 

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© 2007 Humana Press Inc.

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Oldenbourg, R. (2007). Analysis of Microtubule Dynamics by Polarized Light. In: Zhou, J. (eds) Microtubule Protocols. Methods in Molecular Medicine™, vol 137. Humana Press. https://doi.org/10.1007/978-1-59745-442-1_8

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  • DOI: https://doi.org/10.1007/978-1-59745-442-1_8

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-642-9

  • Online ISBN: 978-1-59745-442-1

  • eBook Packages: Springer Protocols

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