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A model based method for the accurate quantification of the
3D structure of fluorescently labelled cellular objects similar
in size to the optical resolution limit is presented. Thismethod
is applied to both simulated confocal images of chromatin
structuresandtoreal confocal data obtainedonaFluorescence
in situ Hybridization (FISH) labelled gene domain. The model
assumes that the object is composed of a small number of
discrete points which are convolved with the microscope
point spread function to give the image. Fitting this model
to image data results in a method to assess object structure
which is accurate, shows a low bias, and does not require user
interventionor thepotentially subjective setting ofathreshold.