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Background subtraction

The image brightness not only originates from fluorescence of the fluorescently labelled molecules of interest. For example detector readout noise, autofluorescence (medium, glass...), and reflected light contribute to the total detected intensity. Therefore the average background value (background measurement in an area outside the cell) should be subtracted from the average pixel value in the bleaching region for each timestep.

The background quantification can be carried out with the confocal operating software (e.g. Zeiss LSM, Leica LCS) or with other image processing software which can handle time series (e.g. the freeware ImageJ).

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contact: Stefan Terjung           last update: 02/06/04