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Study of organic probes and strategies of DNA fluorescent labelling: from basic photochemistry to super-resolution optical microscopy

Grant number: 11/08727-1
Support Opportunities:Scholarships in Brazil - Doctorate (Direct)
Effective date (Start): August 01, 2011
Effective date (End): May 31, 2016
Field of knowledge:Physical Sciences and Mathematics - Chemistry - Physical-Chemistry
Principal Investigator:Marcelo Henrique Gehlen
Grantee:Milena Helmer Lauer
Host Institution: Instituto de Química de São Carlos (IQSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil
Associated scholarship(s):13/09965-9 - Study of polymer systems and organic fluorescent probes by super-resolution optical microscopy, BE.EP.DD


The study of new fluorescente probes is crucial for the application in advanced methods in optical microscopy. In this research, we propose the photophysical study of a set of bisarylated maleic anhydride and maleimide derivatives, synthesized by the Heck-Matsuda reaction. The dynamics of the excited state of these compounds refer to the push-pull effect, where there is a notable charge shift. These kind of compounds are good probes for biological systems, polymers and colloids. The second issue of this research concerns the investigation of an optimal methodology for the DNA sequence-specific fluorescent labelling, using super-resolution fluorescence microscopy. The reactions were carried out utilizing a two-step methodology. For that, a functional group is coupled to the DNA by an enzymatic transfer, then, the fluorophore is coupled to the modified DNA by biorthogonal reactions. A single-molecule assay was developed to count the number of dye molecules covalently bonded to single plasmids. Additionally, Atomic Force Microscopy (AFM) images were recorded to check the morphologies after the labeling reactions. We also propose the study of cyclized maleic anhydride derivatives as DNA intercalation dyes, using super-resolution microscopy. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
LAUER, MILENA HELMER; VRANKEN, CHARLOTTE; DEEN, JOCHEM; FREDERICKX, WOUT; VANDERLINDEN, WILLEM; WAND, NATHANIEL; LEEN, VOLKER; GEHLEN, MARCELO H.; HOFKENS, JOHAN; NEELY, ROBERT K.. Methyltransferase-directed covalent coupling of fluorophores to DNA. CHEMICAL SCIENCE, v. 8, n. 5, p. 3804-3811, . (11/08727-1, 13/09965-9)
LAUER, MILENA HELMER; DREKENER, ROBERTA LOPES; DUARTE CORREIA, CARLOS ROQUE; GEHLEN, MARCELO HENRIQUE. Fluorescence from bisaryl-substituted maleimide derivatives. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. 13, n. 6, p. 859-866, . (11/18215-8, 11/08727-1, 10/12193-0, 11/23832-6)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
LAUER, Milena Helmer. Study of organic probes and strategies for DNA fluorescent labelling: From basic photochemistry to super-resolution optical microscopy. 2016. Doctoral Thesis - Universidade de São Paulo (USP). Instituto de Química de São Carlos (IQSC/BT) São Carlos.

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