Scientific Organisers: Stefanie Reichelt, Alex Sossick, Nick Barry, Alessandro Esposito and Kirti Prakash
The meeting will begin at 1pm BST.
As part of the 'Imaging ONEWORLD' series, the focus of these lectures is on microscopy and image analysis methods and how to apply these to your research. Almost all aspects of imaging such as sample preparation, labelling strategies, experimental workflows, ‘how-to’ image and analyse, as well as facilitating collaborations and inspiring new scientific ideas will be covered. Speakers will be available for questions and answers. The organisers, CRUK CI core facility staff, Gurdon Institute, MRC-LMB, MRC Cancer Unit and NPL will be able to continue the discussion and provide advice on your imaging projects.
Speaker
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Professor Christoph Cremer
Christoph Cremer studied Physics at the University of Munich (LMU) and obtained a Ph.D. (1976) in Biophysics and Genetics, as well as a Dr. med. habil. degree in General Human Genetics and Experimental Cytogenetics (1983) at the University of Freiburg/Germany. Since 1983 he is a Professor for Applied Optics and Information Processing at the University of Heidelberg. From 2011- 2019 he has been a group leader (super-resolution microscopy) at the Institute of Molecular Biology (IMB), Mainz (Germany). In 2013, he was appointed Honorary Professor (Physics) at the University of Mainz (JGU). In addition, he presently is a Research Associate at the two Mainz based Max Planck Institutes for Polymer Research and for Chemistry, respectively. The methodological focus of his research is the development and application of methods of super-resolution fluorescence light microscopy. Contributions to this field include focused nanoscopy, structured illumination, and various types of localization microscopy. As the main application field, these techniques are applied to study the nuclear nanostructure in various cell types and organisms.
Speakers Abstract
The tens of thousands of genes and the related complex biochemical networks allowing their precisely controlled transcription and replication are located in the cell nucleus with a typical size of a few micrometer only. Evidence is accumulating that for these processes, spatial organization is of utmost importance. Hence the structure of the cell nucleus of higher organisms has emerged as a main topic of advanced light microscopy. So far, a variety of methods have been applied, including confocal laser scanning fluorescence microscopy, 4Pi-, STED-, and Localization Microscopy approaches, as well as (laterally) structured illumination microscopy (SIM). Here we discuss application perspectives for nuclear nanostructure analysis of a complementary technique, Spatially Modulated Illumination (SMI). SMI is a widefield based approach to use axially structured illumination patterns to determine the size of small, optically isolated fluorescent objects between about 100 nm and 40 nm diameter with a precision down to the few nm range.
Among the advantages of SMI are simplicity of design, robustness and speed combined with a low illumination intensity. As examples, the SMI analysis of nuclear protein aggregates (Ki 67, transcription factories, replication structures) and of specific small chromatin and gene domains will be highlighted. The SMI instrumentation may be combined with other methods such as SIM or Single Molecule Localization Microscopy (SMLM); numerical simulations indicate that in this way, the axial localization precision may be improved down to the 1 nm range using fluorescence photon statistics suitable for single molecule detection.