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DPG

Regensburg 2004 – wissenschaftliches Programm

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AKB: Biologische Physik

AKB 50: Poster Session "Biological Physics"

AKB 50.94: Poster

Freitag, 12. März 2004, 10:30–13:00, B

DNA in situ hybridization detection by gold conjugated nanoparticles and Atomic Force Microscopy — •Gabriella Teti1, Konstantin Agelopoulos 2, Burkhard Brandt 2, Stefan Thalhammer 1, and Wolfgang Heckl 11Department Geo- und Umweltwissenschaften, Ludwig Maximilians Universitaet-Muenchen — 2Institut klinische Chemie und Laboratoriumsmedizin, Westf*lische Wilhelms-Universitaet Muenster

The localization of specific molecules in biological samples continues to be important for the basic and applied biological research. Immunocytochemistry and in situ hybridization of nucleic acids are key methods. For high resolution localization of specific DNA sequences in situ on biological samples, a study based on the combination of atomic force microscope (AFM) and DNA in situ hybridization technique has been proposed. The DNA probes were labelled with digoxigenin and the detection system was based on antibodies against digoxigenin conjugated with gold particles. In some cases the gold labelling was amplified by a colloidal silver enhancement. Compared to high resolution electron microscopy, AFM generates topographic and three-dimensional images on a nanometre scale in ambient and liquid conditions without destroying the sample morphology. The experimental approach was demonstrated on specific probes against human leukaemia, epidermal growth factor receptor on human metaphase chromosomes and specific oligonucleotides on stretched plasmid DNA. Finally, we suggest potential applications based on our results for high-resolution physical mapping of human genes and disease correlated genes.

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