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, core facility staff from the University of Cambridge, Gurdon Institute, MRC-LMB and the ICR/Royal Marsden Trust are also able to continue the discussion and provide advice on your imaging projects.
Scientific Organisers
Nick Barry
An optical physicist and specialist in light microscopy and head of the Light Microscopy facility at the MRC Laboratory of Molecular Biology, University of Cambridge.
Nabanita Chatterjee
Dr Nabanita Chatterjee is a cell biologist and light microscopist. During her PhD, she identified the role and mechanism of the first glycine transporter in fruit flies required for post translational glycation of sperm tubulin. She presented her doctoral research at a Gordon Conference on Developmental Biology, NH, USA with a Gordon Conference travel award. She received other awards from St John's University, NY, USA besides the Daniel Lilly Scholarship Award for Excellence in Research. She followed the Drosophila into Cold Spring Harbor Laboratory, NY, USA where she looked at the role of transposable elements for epigenetic regulation of aging and neurodegeneration with a focus on ALS. After a brief stint at University of Pittsburgh studying dense core vesicle trafficking in fly larvae, she joined Nikon Microscopes as a Senior Application Scientist with a pan-India customer base. Currently she works at CCI, Gothenburg, as a Light Microscopy Scientific Officer. She is in interested in all things Microscopy and image analysis.
Alessandro Esposito
Dr Alessandro Esposito obtained a PhD in Biophysics in 2006 working at the University of Utrecht and the European Neuroscience Institute in Goettingen for which he was awarded the ‘Sergio Ciani’ award by the Italian Society of Pure and Applied Biophysics. At the University of Cambridge, he then developed novel analytical tools contributing to redefining models of red blood cells homeostasis infected by P. falciparum (malaria). In recognition of his early work, in 2009 Alessandro was awarded a Life Science Interface fellowship by the EPSRC to establish foster the development of heavily multiplexed biochemical imaging. Soon after he moved to the MRC Cancer Unit where he lead the ‘Systems Microscopy initiative’ and retrained in cancer biology. During these years, Alessandro’s work developed into two research streams: i) the study of cellular responses to DNA damage and mutations in signalling pathways and ii) the innovation of biochemical imaging technologies. His team contributed to revealing the vast cell-to-cell variability in stress responses of genetically identical cells, a feature of biological systems that hinder the efficacy of disease management and therapeutic efficacy. Since 2019, Alessandro leads a transdisciplinary research programme at the MRC Cancer Unit in Cambridge devoted to understanding how DNA damage and mutations in KRAS derange homeostatic programmes leading to cancer. His group combines multi-omics data with single-cell biochemical imaging techniques aiming to achieve a deeper understanding of cancer phenotypes during the earliest stages of carcinogenesis, with particular attention to cell-to-cell variability of non-genetic origin and cell-to-cell communication.
Christian Franke
From 2006-2012, Christian studied physics at the University of Würzburg. Afterwards, he joined the group of Prof. Markus Sauer at the Biocenter Würzburg to do his PhD work at the intersection of physics, biology and computer science. During his time at the Sauer Lab he mainly worked in methods development, both hard- and software, for quantitative super-resolution (single-molecule) microscopy. In 2017 he finished his PhD work and started his Postdoctoral research at the Max Planck Institute of Molecular Cell Biology and Genetics Dresden in the department of Prof. Marino Zerial. At MPI-CBG Christian was 'confronted' with complex biological questions regarding the endosomal pathway and its nanoscale representation. Being previously a pure methods developer, now it was all about applying these powerful techniques ‘in the real world’. He developed workflows for correlative super-resolution and electron microscopy imaging of endosomal compartments to match the membrane ultrastructure obtained with EM with the specific nanoscopic localization of fluorescently labelled endosomal markers by dSTORM. Amongst others, his work contributed to the discovery of a novel mechanism of cytosolic escape from endosomal recycling tubules and a first of its kind nanoscale study on the subcellular fate of mRNA, delivered by lipid nanoparticles. In November 2020 he started his own lab as an Assistant Professor of Digitized Experimental Microscopy at the Institute of Applied Optics and Biophysics (IAOB) of the Friedrich Schiller University Jena. Here, he and his team work to combine both the development of high-end tools for microscopy with their application in cell biology and clinical questions.
Kirti Prakash
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.
Fei Xia
Fei is a postdoctoral researcher at Ecole Normale Supérieure in Paris working on computational imaging and optical computing. Fei obtained her Ph.D. from Cornell University in 2021, with thesis work on deep tissue imaging tool development with short-wave infrared light and adaptive optics. Her current research interest is in computational and hardware tool development for real-world applications and fundamental science, particularly in biomedicine.
Advanced light microscopy for studying the structure and dynamics of the cardiovascular system
The cardiovascular network is crucial for all body functions. Since functional or anatomical abnormalities in this network are associated with a wide range of patho-physiologies, studying these aspects is of fundamental and clinical importance. In my talk I will discuss our recent efforts to study the structure and dynamics of different components of the cardiovascular network using advanced optical imaging. I will focus on two techniques that have relevant spatio-temporal scales, in the range of subcellular/millisecond resolutions. The first is light-sheet microscopy, which employs an orthogonal excitation-emission geometry that enables fast imaging, and is becoming a standard tool for probing large specimens and fast biological processes. The second technique utilizes optical harmonic imaging signals, which are signals that are generated by different endogenous tissue components. Harmonic imaging is a non-linear “label-free” imaging method that obviates the need for fluorescent or chemical contrast agents, and is thus potentially useful for simple and straight-forward analysis of tissue structures. I will demonstrate how we used light-sheet microscopy separately from, or in combination with, harmonic imaging, to learn about the cardiovascular network in zebrafish embryos or cerebral vessels in the mouse brain, using tissue transparization. I will also present customized image analysis workflows that we developed in order to analyze the datasets, and will discuss the opportunities that the ensemble of these novel techniques offer for research in the cardiovascular field and more broadly for understanding biological and physiological processes as they take place.
