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.
University of Cambridge
University of Cambridge
Alex heads the Imaging Facility at the Gurdon Institute, which includes a variety of microscopy techniques including confocal, high throughput and deconvolution. He is keen to raise the level of microscopy understanding and application, and runs and takes part in various microscopy courses.
CRUK Cambridge Institute
Stefanie Reichelt, PhD is head of the light microscopy facility at the CRUK Cambridge Institute. The core provides state-of-the-art imaging resources, training courses for scientists and students and develop new imaging systems as well as user-friendly analysis and acquisition tools for specific research applications. Stefanie teaches academically at Cambridge University, in scientific workshops and out-reach events. She is also founder of the well-known Plymouth Advanced Microscopy Course and the Centre for Mathematical Imaging in Healthcare (CIMH)
MRC Cancer Cell Unit
Alessandro’s major focus is to develop new methods for probing and mapping the biochemical events that underlie cellular systems using fluorescence microscopy, with the overall aim of better understanding the fundamental principles that govern tumour suppressive mechanisms and their perturbation in cancer. In his career, Alessandro has developed several technologies (e.g. high throughput FLIM, solid state detectors and imaging specotrpolarimetry) and methods of analysis for quantitative biochemical imaging (FLIM, spectral imaging and anisotropy imaging). Since 2009, he has focused on strategies to enhance the biochemical resolution of fluorescence microscopy and multiplexing several biochemical reactions within the cell with the scope to correlate complex biochemical signatures to cell fate decision and homeostatic control.
MRC-Laboratory of Molecular Biology
Institute for Cancer Research (ICR) and Royal Marsden Trust
Kirti Prakash is a computer scientist by training (Bachelors and Masters degree) but a biologist at heart (PhD degree). Kirti aspires to be an inventor and develop new imaging tools for cell biology and neuroscience. Kirti did his Masters in Computer Science from Aalto University (Finland) and PhD in Biology from Heidelberg University (Germany). During his PhD, he developed a new method to image DNA which led to the first high-resolution images of the epigenetic landscape of meiotic chromosomes and mechanisms behind chromosome condensation. The doctoral research earned him several awards including Springer Best PhD Thesis Prize. After his PhD, he did a couple of postdocs at Carnegie Institution for Science (USA) and University of Cambridge (UK). The primary highlights of his research here were laser-free superresolution microscopy and development of a high-content imaging pipeline to quantify single-cell gene expression. Formerly at the National Physical Laboratory (NPL), and currently working at the Institute for Cancer Research (ICR) and Royal Marsden Trust, he is working on microscope development and image analysis.
The accurate detection and repair of DNA damage is crucial for genome stability in all organisms. Despite extensive characterization of DNA repair pathways using genetics and biochemical assays, it remains unclear how repair proteins perform their function within the cellular environment. I will present our developments of microfluidic imaging techniques to investigate DNA repair and mutagenesis in individual living bacterial cells. By combining super-resolution localisation microscopy and single-molecule tracking, we were able to directly follow the movement and activity of DNA repair enzymes and regulatory proteins inside cells. This approach provided new insight into how DNA repair proteins identify lesion sites within the crowded cellular environment.
A key advantage of single-cell and single-molecule techniques is their ability to resolve biological heterogeneity and dynamic behaviour without ensemble averaging. Using these approaches, we found that stochasticity, or "noise", can play important roles in the function of the DNA repair system. By modulating mutation rates, noise in DNA repair can create a cell subpopulation that acts as a source of genetic diversity during adaptation to stress conditions.
Group Leader Department of Biochemistry, Oxford UniversityStephan Uphoff is a group leader at the Department of Biochemistry of Oxford University and a tutorial fellow at New College. He studied physics in Göttingen and specialised in single-molecule biophysics for his MSc and DPhil at Oxford. He moved to Harvard Medical School for postdoctoral research in the field of systems biology and returned to Oxford to join the Biochemistry Department as a Wellcome Trust fellow. Stephan established his independent group in 2017. He is the recipient of the Colworth Medal of the Biochemical Society, the Lister Institute research prize, and the Wellcome-Beit prize. His multi-disciplinary research group develops and applies microscopy techniques to study how bacteria adapt to stress conditions, with a particular focus on the mechanisms and functions of DNA repair and mutagenesis.