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  • 1
    Publication Date: 2020-01-28
    Description: The events of meiosis I in a living spermatocyte obtained from the testis of a crane-fly larva are recorded in this time-lapse sequence beginning at diakinesis through telophase to the near completion of cytokinesis following meiosis I.
    Keywords: Meiosis I ; Spermatocyte ; Microtubules ; Kinetochore microtubules ; Polarized light microscopy ; LC-PolScope
    Repository Name: Woods Hole Open Access Server
    Type: Moving Image
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  • 2
    Publication Date: 2020-01-28
    Description: The events of meiosis II in two living spermatocytes obtained from the testis of a crane-fly larva are recorded in this time-lapse sequence beginning at prophase II through telophase II to the near completion of cytokinesis following meiosis II.
    Keywords: Meiosis II ; Spermatocyte ; Microtubules ; Kinetochore microtubules ; Polarized light microscopy ; LC-PolScope
    Repository Name: Woods Hole Open Access Server
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  • 3
    Publication Date: 2022-05-25
    Description: Author Posting. © Microscopy Society of America, 2007. This article is posted here by permission of Cambridge University Press for personal use, not for redistribution. The definitive version was published in Microscopy and Microanalysis 13 Suppl. 2 (2007): 10-11, doi:10.1017/S1431927607075186.
    Description: Differential interference contrast (DIC) microscopy is widely used to observe structure and motion in unstained, transparent living cells and isolated organelles, producing a monochromatic shadowcast image of optical phase gradient. Polarized light microscopy (Pol) reveals structural anisotropy due to form birefringence, intrinsic birefringence, stress birefringence, etc. DIC and Pol complement each other as, for example, in a live dividing cell, the DIC image will clearly show the chromosomes while the Pol image will depict the distribution of the birefringent microtubules in the spindle. Both methods, however, have the same shortcomings: they require the proper orientation of a specimen in relation to the optical system in order to achieve best results.
    Repository Name: Woods Hole Open Access Server
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  • 4
    Publication Date: 2022-05-25
    Description: Author Posting. © American Society for Cell Biology, 2004. This article is posted here by permission of American Society for Cell Biology for personal use, not for redistribution. The definitive version was published in Molecular Biology of the Cell 15 (2004): 5346-5355, doi:10.1091/mbc.E04-06-0524.
    Description: To test the "traction fiber" model for metaphase positioning of bivalents during meiosis, kinetochore fibers of maloriented bivalents, induced during recovery from cold arrest, were analyzed with a liquid crystal polarizing microscope. The measured birefringence retardation of kinetochore fibers is proportional to the number of microtubules in a fiber. Five of the 11 maloriented bivalents analyzed exhibited bipolar malorientations that had at least four times more kinetochore microtubules to one pole than to the other pole, and two had microtubules directed to only one pole. Yet all maloriented bivalents had positions at or near the spindle equator. The traction fiber model predicts such maloriented bivalents should be positioned closer to the pole with more kinetochore microtubules. A metaphase position at the spindle equator, according to the model, requires equal numbers of kinetochore microtubules to both poles. Data from polarizing microscope images were not in accord with those predictions, leading to the conclusion that other factors, in addition to traction forces, must be involved in metaphase positioning in crane-fly spermatocytes. Although the identity of additional factors has not been established, one possibility is that polar ejection forces operate to exert away-from-the-pole forces that could counteract pole-directed traction forces. Another is that kinetochores are "smart," meaning they embody a position-sensitive mechanism that controls their activity.
    Description: J.R.L. is supported by grant MCB-0235934 from the National Science Foundation. R.O. is supported by grants GM49210 from the National Institute of General Medical Sciences and EB002045 from the National Institute of Biomedical Imaging and Bioengineering.
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  • 5
    Publication Date: 2022-05-25
    Description: Author Posting. © American Society for Cell Biology, 2007. This article is posted here by permission of American Society for Cell Biology for personal use, not for redistribution. The definitive version was published in Molecular Biology of the Cell 18 (2007): 1645-1656, doi:10.1091/mbc.E06-10-0963.
    Description: This study investigated the basis of meiosis II nondisjunction. Cold arrest induced a fraction of meiosis II crane fly spermatocytes to form (n + 1) and (n – 1) daughters during recovery. Live-cell liquid crystal polarized light microscope imaging showed nondisjunction was caused by chromosome malorientation. Whereas amphitely (sister kinetochore fibers to opposite poles) is normal, cold recovery induced anaphase syntely (sister fibers to the same pole) and merotely (fibers to both poles from 1 kinetochore). Maloriented chromosomes had stable metaphase positions near the equator or between the equator and a pole. Syntelics were at the spindle periphery at metaphase; their sisters disconnected at anaphase and moved all the way to a centrosome, as their strongly birefringent kinetochore fibers shortened. The kinetochore fibers of merotelics shortened little if any during anaphase, making anaphase lag common. If one fiber of a merotelic was more birefringent than the other, the less birefringent fiber lengthened with anaphase spindle elongation, often permitting inclusion of merotelics in a daughter nucleus. Meroamphitely (near amphitely but with some merotely) caused sisters to move in opposite directions. In contrast, syntely and merosyntely (near syntely but with some merotely) resulted in nondisjunction. Anaphase malorientations were more frequent after longer arrests, with particularly long arrests required to induce syntely and merosyntely.
    Description: This study was supported by National Science Foundation grant MCB-0235934 (to J.L.) and grants GM 49210 from the National Institute of General Medical Sciences and EB002045 from the National Institutes of Biomedical Imaging and Bioengineering (to R.O.).
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