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Appendix 4 Determination of contamination level in IF and FISH tests

United Kingdom

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  1. 1..

    Count the mean number of typical fluorescent cells per field of view (c)

  2. 2..

    Calculate the number of typical fluorescent cells per microscope slide window (C)

    C = c × S/s

    whereS=surface area of window of multispot slide, and
    s=surface area of objective field.
    s = πi2/4G2K2 wherei=field coefficient (varies from 8 to 24 depending upon ocular type)
    K=tube coefficient (1 or 1,25)
    G=magnification of objective (100x, 40x etc.).
  3. 3..

    Calculate the number of typical fluorescent cells per ml of resuspended pellet (N)

    N = C × 1 000/y × F

    wherey=volume of resuspended pellet on each window, and
    F=dilution factor of resuspended pellet.