This book will serve as an introduction to microscopy and biomedical imaging methods, with a focus 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 noninvasive 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, which 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 that 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 correla-tion 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 to graduate and upper-level undergraduate students and industry researchers in the fields of cell biology, microbiology, microscopy, and medical imaging.
Chapter 1. General introduction to imaging the Plasma membrane
Chapter 2. How to map membrane proteins at the single-molecule level
Chapter 3. Measuring diffusion of membrane biomolecules using fluorescence microscopy
Chapter 4. Detecting membrane protein dimers and oligomers by FRET
Chapter 5. Imaging membrane biophysical properties
Chapter 6. Conclusions and future perspectives