Method for spatial high-resolution investigation of a structure of a sample marked with a substance

摘要

The method for spatial high-resolution examination of a biological structure (1) of a sample marked with a substance (3), comprises converting the substance from a first state such as a non-fluorescent state into a second state such as a fluorescent state, where the first and second states differ from each other in a photo-physical property, initially binding a protein tag (2) on the structure for marking the structure or expressing the protein tag together with the structure as fusion protein, and then non-covalently binding the substance by the tag. The tag has fluorogen activating protein. The method for spatial high-resolution examination of a biological structure (1) of a sample marked with a substance (3), comprises converting the substance from a first state such as a non-fluorescent state into a second state such as a fluorescent state, where the first and second states differ from each other in a photo-physical property, initially binding a protein tag (2) on the structure for marking the structure or expressing the protein tag together with the structure as fusion protein, and then non-covalently binding the substance by the tag. The tag has fluorogen activating protein. The substance is only switchable or convertible from the first state into the second state in a state unbound with the fluorogen activating protein. A bleached molecule of the substance is substituted by an unbleached molecule of the substance during the state unbound with the fluorogen activating protein. The tag has a part binding the structure and a part binding the substance. The part binding the structure non-covalently binds to the part binding the substance. An adapter protein is realized between the part binding the structure and the part binding the substance. The photo-physical properties of the substance are influenced by providing or changing local environmental conditions. The change of the photo-physical properties of the substance takes place by binding or the type of binding to the fluorogen activating protein. The fluorogen activating protein-type is selected or adapted in dependent of the examination technique or property of the examination technique.