Department of Biological and Medical Sciences

Methods

  • Protocols from IPNC member labs

    Nuclei stains for live cell imaging (confocal)

    Ethidium bromide

    Mix 1:1 with 1x PBS, immerse plant tissue (roots, leaf sections, anthers) for 1-5mins

    Excite with 514nm, sample fluorescence with 530-600nm

    DRAQ5

    from Biostatus

    test in BY-2 cells

    works best with Triton but have used with BY-2 cells without Triton

    1. For BY-2's without Triton, add 1ul neat DRAQ5 to 200ul BY-2 cells and incubate for 20-40min
    2. With Triton: To 500 μl cells add 1 μl 20% Triton X-100 (1 μl Triton X-100 in 5 μl dH2O) and 0.2 μl of 5 mM DRAQ5 for a working concentration of 10 μM; incubate 1-2min before imaging

    Excitation with any laser from 488nm upwards; capture excitation in LP650.