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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 191 (1996), S. 215-219 
    ISSN: 1615-6102
    Keywords: Laser scanning confocal microscopy ; Multiple cell layers ; Plant microtubules ; Plant microfilaments ; Roots ; Tissue clearing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A protocol was developed to observe plant microtubules and actin microfilaments in large tissue samples without physical sectioning. Rye (Secale cereale L. cv. Rymin) root tip pieces from two-day-old seedlings were fixed and processed for immunolabeling. Incubation times of 24–48 h were required to insure adequate penetration of fixatives, antibodies, and washing buffers. Clearing of the tissue with methyl salicylate reduced background auto-fluorescence that would otherwise interfere with the resolution of cytoskeletal structures. Microtubules or microfilaments in 5–7 cell layers were visualized using the optical-sectioning capability of laser scanning confocal microscopy (LSCM) and projected as three-dimensional images. The three-dimensional character of the cytoskeletal elements is retained when viewing stained cells of intact tissue. The net-like character of a microfilament array radiating out from a single point into the cytoplasm is maintained when the cells are stained in intact root tip pieces and imaging is accomplished in situ.
    Type of Medium: Electronic Resource
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