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Using Fluorescence Microscopy to Study Mitosis

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The Mitotic Spindle

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1413))

Abstract

Fluorescence microscopy is one of the most important approaches in the cell biologist’s toolbox for studying the mitotic spindle. In fact, many of the key insights into our understanding of mitosis have been enabled by the visualization of mitotic processes using fluorescence microscopy. Here, we summarize some of the important considerations for imaging mitosis using fluorescence microscopy. Because light can damage live cells, we emphasize the importance of minimizing cellular damage while obtaining informative images.

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References

  1. Poser I, Sarov M, Hutchins JRA, Heriche J-K, Toyoda Y, Pozniakovsky A, Weigl D, Nitsche A, Hegemann B, Bird AW, Pelletier L, Kittler R, Hua S, Naumann R, Augsburg M, Sykora MM, Hofemeister H, Zhang Y, Nasmyth K, White KP, Dietzel S, Mechtler K, Durbin R, Stewart AF, Peters J-M, Buchholz F, Hyman AA (2008) BAC transgeneOmics: a high-throughput method for exploration of protein function in mammals. Nat Methods 5:409–415

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Munoz IM, Szyniarowski P, Toth R, Rouse J, Lachaud C (2014) Improved genome editing in human cell lines using CRISPR method. Plos One 9:e109752

    Article  PubMed  PubMed Central  Google Scholar 

  3. Pawley JB (2006) Handbook of biological confocal microscopy, 3rd edn. Springer, New York, NY

    Google Scholar 

  4. Wadsworth P (2010) Studying mitosis in cultured mammalian cells. In: Goldman RD, Swedlow JR, Spector DL (eds) Live cell imaging a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp 571–582

    Google Scholar 

  5. Swedlow JR (2010) In vivo imaging of mammalian cells. In: Goldman RD, Swedlow JR, Spector DL (eds) Live cell imaging a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp 317–331

    Google Scholar 

  6. Goodson HL, Dzurisn JS, Wadsworth P (2010) Methods for expressing and analyzing GFP-tubulin and GFP-microtubule-associated proteins. In: Goldman RD, Swedlow JR, Spector DL (eds) Live cell imaging: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp 605–622

    Google Scholar 

  7. Wadsworth P, Rusan NM, Tulu US, Fagerstrom C (2005) Stable expression of fluorescently tagged proteins for studies of mitosis in mammalian cells. Nat Methods 2:981–987

    Article  CAS  PubMed  Google Scholar 

  8. Maddox P, Moree B, Canman JC, Salmon ED (2003) Spinning disk confocal microscope system for rapid high-resolution, multimode, fluorescence speckle microscopy and green fluorescent protein imaging in living cells. Methods Enzymol 360:597–617

    Article  PubMed  Google Scholar 

  9. Rose GG, Pomerat CM, Shindler TO, Trunnel JB (1958) A cellophane strip technique for culturing tissue in multipurpose culture chambers. J Biophys Biochem Cytol 4:761–764

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Waters JC (2009) Accuracy and precision in quantitative fluorescence microscopy. J Cell Biol 185:1135–1148

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Swedlow JR, Sedat JW, Agard DA (1997) Deconvolution in optical microscopy. In: Jansson PA (ed) Deconvolution of images and spectra. Academic, New York, pp 284–309

    Google Scholar 

  12. Rusan NM, Wadsworth P (2005) Centrosome fragments and microtubules are released and transported asymmetrically away from division plane in anaphase. J Cell Biol 168:21–28

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Winter PW, Shroff H (2014) Faster fluorescence microscopy: advances in high speed biological imaging. Curr Opin Chem Biol 20:46–53

    Article  CAS  PubMed  Google Scholar 

  14. Lee W-L, Oberle JR, Cooper JA (2003) The role of the lissencephaly protein Pac1 during nuclear migration in budding yeast. J Cell Biol 160:355–364

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Rizzo MA, Davidson MW, Piston DW (2010) Fluorescence protein tracking and detection. In: Goldman RD, Swedlow JR, Spector DL (eds) Live cell imaging a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp 3–34

    Google Scholar 

  16. Magidson V, O’Connell CB, Loncarek J, Paul R, Mogilner A, Khodjakov A (2011) The spatial arrangement of chromosomes during prometaphase facilitates spindle assembly. Cell 146:555–567

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Correspondence to Patricia Wadsworth .

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Balchand, S.K., Mann, B.J., Wadsworth, P. (2016). Using Fluorescence Microscopy to Study Mitosis. In: Chang, P., Ohi, R. (eds) The Mitotic Spindle. Methods in Molecular Biology, vol 1413. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3542-0_1

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  • DOI: https://doi.org/10.1007/978-1-4939-3542-0_1

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3540-6

  • Online ISBN: 978-1-4939-3542-0

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