Two-color intranuclear distance measurements of gene regions in human lymphocytes

S. Fenz; H. Mathée; G. Kreth; D. Baddeley; Y. Weiland; J. Schwarz-Finstlerle; C. G. Cremer; U. J. Birk

doi:10.1117/12.727506

12 July 2007 Two-color intranuclear distance measurements of gene regions in human lymphocytes

S. Fenz, H. Mathée, G. Kreth, D. Baddeley, Y. Weiland, J. Schwarz-Finstlerle, C. G. Cremer, U. J. Birk

Author Affiliations +

Proceedings Volume 6630, Confocal, Multiphoton, and Nonlinear Microscopic Imaging III; 663002 (2007) https://doi.org/10.1117/12.727506
Event: European Conferences on Biomedical Optics, 2007, Munich, Germany

Abstract

The method of Spectral Precision Distance Microscopy (SPDM) has been used to determine distances between two FISH (Fluorescence-in situ-Hybridization)-labeled gene regions on chromosome 9. To this end we applied methods to correct for chromatic aberrations of the microscope optics alone and also of the sample induced aberrations due to mismatch of the refractive indices. Using a confocal microscope and a threshold based position determination algorithm, positions could be measured with an accuracy of about 65 nm inside of fixed cell nuclei. Distances obtained from the measurements have been verified using a 3D computer model of the cell nucleus. In principle, this SPDM approach could be combined with novel fluorescence microscopes to obtain structural information well below the optical resolution. At present the precision limit of the distance measurements is set by variations of the refractive index throughout the specimens.

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S. Fenz, H. Mathée, G. Kreth, D. Baddeley, Y. Weiland, J. Schwarz-Finstlerle, C. G. Cremer, and U. J. Birk "Two-color intranuclear distance measurements of gene regions in human lymphocytes", Proc. SPIE 6630, Confocal, Multiphoton, and Nonlinear Microscopic Imaging III, 663002 (12 July 2007); https://doi.org/10.1117/12.727506

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