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This book will serve as an introduction to microscopy and
biomedical imaging methods, focussed on the study of the
distributions and dynamics of molecules on the cell surface. It
will provide readers with an in-depth understanding of how modern
microscopy methodology can be used to understand the organisation
of cell membrane systems and how experiments can be designed around
these methodologies. There are numerous methods employed to
understand cell membrane organisation, but foremost among them are
microscopy methods which can map the distributions of molecules on
the cell surface and even map the biophysical properties of
membranes themselves. Fluorescence microscopy has been especially
widely used due to its specificity and relatively non-invasive
nature, allowing live-cell imaging. However, the recent advance of
super-resolution fluorescence microscopy has broken the previous
resolution limit for this type of microscopy has been an important
advancement in the field. Atomic force microscopy and electron
microscopy have also been deployed to learn about membrane
organisation and properties. Each chapter in this volume will be
themed around measuring a particular property of cell membranes. In
each case the authors examine the range of methodology applicable
to the task, comparing the advantages and disadvantages of each
one, and will also provide an overview of important discoveries
which have been made using the methodology being discussed. The
chapters will cover: · Measuring membrane protein distributions
using single-molecule localisation microscopy (SMLM) · Measuring
membrane protein dynamics and diffusion using fluorescence
correlation spectroscopy (FCS) · Mapping membrane lipid backing
using environmentally sensitive fluorescence probes · Mapping
membrane thickness and rigidity using atomic force microscopy ·
Mapping membrane proteins and the cytoskeleton using electron
microscopy This book will be a valuable resource be to graduate and
upper-level undergraduate students and industry researchers in the
fields of cell biology, microbiology, microscopy, and medical
imaging.
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