Quantitative phase imaging microscopy for label-free, high-content imaging

点击翻译按钮获取中文摘要

Optical microscopy is a routine tool in biomedical research, allowing us to view the inner workings of cells and how cells work together to form tissues. Being able to see the movement of cells helps us to understand important processes in healthy cells and tissues and what goes wrong in disease.Cells are inherently transparent and difficult to image in their native state. Typically, we add fluorescent markers to enable visualisation of cellular substructures or specific proteins. While such fluorescent probes result in excellent high contrast images, both the introduction and imaging of the probe can be detrimental to cell viability and disturb the observed process. The binding of a bulky fluorescent molecule to a protein of interest, for example, can affect the behaviour of that molecule, while fluorescence imaging produces free radicals which are harmful to the cells. In some cells, such as patient-derived cells or other sensitive cell types, it may not be possible to add fluorescent markers at all, making fluorescence imaging impossible.Light transmitted through transparent cells undergoes a phase shift, by interfering this light with a reference beam, we can generate holograms. Spatial information in the form of cellular density can be extracted from these holograms using carefully designed algorithms. The images generated from these holograms resemble those achieved from fluorescently labelled cells, supplying high contrast images without requiring fluorescent markers. This technique, known as quantitative phase imaging, allows us to image cells in their native state with minimal perturbation.This application requests funds to purchase a quantitative phase imaging microscope for minimally invasive cellular imaging. The requested microscope will come with fully automated acquisition and analysis, enabling us to image up to 96 samples in parallel and extract information on the proliferation, movement and shape of cells automatically in an objective manner. This microscope will therefore support cell-based screening for a range of discovery and translational research projects to understand and develop treatments for rare genetic diseases, bacterial infections and cancer.