Single-molecule and super-resolution
An important part of our group works on the development of single-molecule and super resolution methods. In particular, we develop single-molecule localization microscopy using adaptive optics, implement variants of structured illumination microscopy, and work on the development of multi-focus microscopies. These implementations are combined with other methods such as microfluidics or DNA origami.
3D-single-molecule localization microscopy
We develop several flavors of single-molecule localization microscopy (SMLM). These include
Principles of super-resolution microscopy
Components of our basic SMLM microscope
Super-resolution imaging of bacteria in a micro-fluidics device.
Cattoni* DI, Fiche* JB, Valeri A, Mignot T, Nöllmann M.
PLoS ONE 8(10):e76268, Oct 2013. [PDF]
Single-molecule super-resolution imaging in bacteria
Diego I. Cattoni, Jean-Bernard Fiche, Marcelo Nöllmann
Current Opinion in Microbiology, 15(6):758-63, Dec 2012. [PDF]
Nanometer resolved single-molecule colocalization of nuclear factors by two-color super resolution microscopy imaging
Mariya Georgieva; Diego I Cattoni; Jean-Bernard Fiche; Thibaut Mutin; Delphine Chamousset; Marcelo Nollmann. Methods pii: S1046-2023(16)30063-9. 1 Apr 2016 . [PDF]
We develop new microscopies based on multi-focus microscopy (Abrahamsson, 2013). In particular, we focus on the development and construction of MFG gratings in collaboration with the LAAS (CNRS, Toulouse). Recently, we developed a 3D super-resolution microscopy method that enables deep imaging in cells based on a combination of Multifocus Microscopy and PSF engineering. We demonstrated the performance of the method by imaging the nuclear envelope of eukaryotic cells reaching a depth of field of ~ 4μm. Also, we improved the efficiency of transmission of MFGs by using multi-phase engraving.
Astigmatic multifocus microscopy enables deep 3D super-resolved imaging.
Laura Oudjedi, Jean-Bernard Fiche, Sara Abrahamsson, Laurent Mazenq, Aurélie Lecestre, Pierre-François Calmon, Aline Cerf, and Marcelo Nöllmann
Biomedical Optics Express, 2016, in press. [PDF]
Perfect color Multifocus Microscopy (MFM) with increased sensitivity
Sara Abrahamsson, Rob Ilic, Jan Wisniewski, Brian Mehl, Liya Yu, Lei Chen, Marcelo Davanco, Laura Oudjedi, Jean-Bernard Fiche, Bassam Hajj, Xin Jin, Joan Pulupa, Christine Cho, Mustafa Mir, Xavier Darzacq, Marcelo Nollmann, Maxime Dahan, Carl Wu, Timothy Lionett, James . Liddle and Cornelia I. Bargmann
Biomedical Optics Express, 2016, in press
Magnetic Tweezers coupled to single-molecule fluorescence detection
We developed a standard magnetic tweezers coupled to TIRF illumination and single-molecule fluorescence detection to monitor the activity, mechanical properties, and position of fluorescently-labelled proteins in real-time. The setup has been used for the study of SpoIIIE (Thakur, submitted) and the mechanism of looping of insulator proteins.
The fluorescence properties and binding mechanism of SYTOX green, a bright, low photo-damage DNA intercalating agent.
Thakur, S,Cattoni DI, Nollmann M
European Biophysical Journal, July 2015, 44(5):337-48. [PDF]
Direct observation of the translocation mechanism of transcription termination factor Rho
Gocheva V, Le Gall A, Boudvillain M, Margeat E, Nollmann M
Nucleic Acids Research, 2015 Feb 27; 43(4):2367-77. [PDF]
Constructing a magnetic tweezers to monitor RNA translocation at the single-molecule level.
Salas D, Gocheva V, Nöllmann M.
Methods Mol Biol. 1259:257-73. Jan 2015. [PDF]