Model based precision structural measurements on barely resolved objects.
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. This method is applied to both simulated confocal images of chromatin structures and to real confocal data obtained on a Fluorescence 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 intervention or the potentially subjective setting of a threshold.
Baddeley
D
D
Kirchhoff Institut für Physik, Universität Heidelberg, Im Neuenheimer Feld 227, D-69120 Heidelberg, Germany. d.baddeley@auckland.ac.nz
Weiland
Y
Y
Batram
C
C
Birk
U
U
Cremer
C
C
D002467:Cell Nucleus / Q000648:ultrastructure; D002843:Chromatin / Q000648:ultrastructure*; D003198:Computer Simulation; D015342:DNA Probes; D005456:Fluorescent Dyes; D005796:Genes; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D008214:Lymphocytes / Q000648:ultrastructure; D018613:Microscopy, Confocal / Q000379:methods; D008954:Models, Biological; D008968:Molecular Conformation; D011218:Prader-Willi Syndrome / Q000235:genetics; D012680:Sensitivity and Specificity; D055517:snRNP Core Proteins / Q000235:genetics / Q000648:ultrastructure
D002843:Chromatin; D015342:DNA Probes; D005456:Fluorescent Dyes; C526417:SNRPN protein, human; D055517:snRNP Core Proteins