Abstract
An overview is given on experiments in single molecule biophysics, cell biology and biomedicine, where the extreme sensitivity of fluorescence and/or the extreme specificity, particularly of protein molecules fluorescence labelled with green fluorescent protein, is used to study the basic aspects of life. The report starts with applications on the biophysics of DNA, shows that it behaves as a harmonic spring and shows how a single molecule of the DNA restriction endonuclease, EcoRI cuts a single DNA molecule. Then the comet assay and variants of it are used to show that the DNA repair capacity of human cells remains surprisingly constant up to a high age. Antibiotic and anti-psychotic drugs as well as anticancer drugs may cause, in combination with UV, DNA damages. The different human chromosomes show a differential DNA repair capacity. A variant of the “Comet assay,” the immune fluorescent comet assay allows to detect single DNA breaks in whole cell nuclei. Experiments using a laser microbeam show that in humans, DNA repair is accomplished by non-homologous end joining, a comparably erroneous DNA repair pathway, which may be switched to the more accurate homologous recombination repair pathway. Using a variant of optical tweezers (erythrocyte-mediated force application, EMFA) it is shown that the fibroblast scars generated after a heart infarction may be made transparent for calcium waves and be thus less hazardous of generating deadly “fibrillations.” Finally, it is shown how EMFA and fluorescence-labelled nitric oxide may be used to find new, blood pressure relaxing drugs.
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Greulich, K.O., Grigaravicius, P., Monajembashi, S. (2012). Single DNA Molecule Typing, Heart Beating, DNA Repair and Ageing: The Contribution of Fluorescence Techniques. In: Geddes, C. (eds) Reviews in Fluorescence 2010. Reviews in Fluorescence, vol 2010. Springer, New York, NY. https://doi.org/10.1007/978-1-4419-9828-6_9
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