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New Methods and Sensors for Membrane and Cell Volume Research

1st Edition - November 29, 2021

Editors: Michael Model, Irena Levitan

Hardback ISBN:

9 7 8 - 0 - 3 2 3 - 9 1 1 1 4 - 6

eBook ISBN:

9 7 8 - 0 - 3 2 3 - 9 1 1 1 5 - 3

New Methods and Sensors for Membrane and Cell Volume Research, Volume 88 provides an overview of novel experimental approaches to study both the cell membrane and the… Read more

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New Methods and Sensors for Membrane and Cell Volume Research, Volume 88 provides an overview of novel experimental approaches to study both the cell membrane and the under-membrane space – the cytosol, which have lately began drawing renewed attention. The book's overall emphasis is on fluorescent and FRET-based sensors, however, other optical (such as variants of transmission microscopy) and non-optical methods (neutron scattering and mass spectrometry) also have dedicated chapters. This volume provides a rare review of experimental approaches to study intracellular phase transitions, as well as anion channels, membrane tension and dynamics, and other topics of intense current interest.

Cover
Title page
Table of Contents
Copyright
Contributors
On, in, and under membrane
References
Chapter One: Fluorescence-based sensing of the bioenergetic and physicochemical status of the cell
Abstract
1: Introduction
2: Currently available fluorescent molecules
3: Labeling of (macro)molecules of interest
4: Sensors and methods of detection
5: Tracking of molecular and global changes
6: Microscopy techniques
7: A map to navigate the fluorescent sea
8: Conclusions
Acknowledgments
References
Chapter Two: Current methods for studying intracellular liquid-liquid phase separation
Abstract
1: Characteristics of liquid-liquid phase separation
2: Liquid-liquid phase separation in biology
3: In vivo and in vitro methods of liquid-liquid phase separation detection
4: Computational methods for liquid-liquid phase separation prediction and modeling
5: Databases on liquid-liquid phase separation and intrinsically disordered proteins
6: Summary
Acknowledgments
References
Chapter Three: Investigating molecular crowding during cell division and hyperosmotic stress in budding yeast with FRET
Abstract
1: Introduction
2: Results and discussion
3: Conclusion and discussion
4: Materials and methods
References
Chapter Four: The expanding toolbox to study the LRRC8-formed volume-regulated anion channel VRAC
Abstract
1: Introduction
2: Animal models reveal the physiological functions of VRAC
3: Electrophysiological measurement of VRAC-mediated currents
4: Measurement of transported substrate
5: Measurement of cell volume
6: Monitoring VRAC activity by optical imaging
References
Chapter Five: Studying cell volume beyond cell volume
Abstract
1: Introduction
2: CV measurement by dye exclusion
3: MC measurement by TIE/TTD
4: MC at work
5: Conclusions
Acknowledgments
Appendix
References
Chapter Six: Membrane tension
Abstract
References
Chapter Seven: Methods for assessment of membrane protrusion dynamics
Abstract
1: Introduction
2: Approaches to assess physical changes in cell morphodynamics
3: Approaches to theoretical modeling of cell protrusion activity
4: Biochemical assessment of membrane protrusion dynamics
5: Conclusion
Acknowledgment
References
Chapter Eight: Evaluating membrane structure by Laurdan imaging: Disruption of lipid packing by oxidized lipids
Abstract
1: Introduction
2: Basic principles of Laurdan fluorescence
3: Laurdan two photon imaging: Visualizing domains in membrane vesicles
4: Laurdan two photon imaging: Visualizing membrane domains in living cells
5: Impact of cholesterol depletion on Laurdan GP values
6: Opposite effects of low-density lipoproteins (LDL) and oxidized low-density lipoproteins (oxLDL) on the lipid order of endothelial cells
7: Disruption of lipid packing of endothelial membrane by oxidized lipids: Oxysterols and oxidized phospholipids
8: Challenges and new directions
Acknowledgments
References
Chapter Nine: Fluorescence sensors for imaging membrane lipid domains and cholesterol
Abstract
1: Introduction
2: Fluorescent sensors for imaging lipid domains in situ
3: Sensing transbilayer topography of lipids, cholesterol, and lipid domains
4: Sensing membrane polarity
5: Super-resolution microscopy triggers the development of new sensors
6: Conclusions and future directions
References
Chapter Ten: Mass spectrometry-based lipid analysis and imaging
Abstract
1: Introduction to matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI)
2: MALDI MSI lipid imaging workflow
3: MALDI MSI data analysis
4: Lipid identification in MSI
5: Applications of MALDI MSI in Niemann-Pick type C
6: Current perspective
References
Chapter Eleven: Deciphering lipid transfer between and within membranes with time-resolved small-angle neutron scattering
Abstract
1: Introduction
2: Small angle neutron scattering (SANS) in the study of membranes
3: Kinetic and thermodynamic characteristics of the transport of lipids and sterols between and within membranes obtained from TR-SANS measurements
4: Transport behavior of lipids and sterols in membranes
5: Current and future perspectives
Acknowledgments
References

Michael Model

Michael Model got an M.S. in materials science from Russia and continued his education at the University of Michigan, where he received a Ph.D. in biophysics in 1995. Since 2004, he has been working at Kent State University, first as a microscopy facility manager and currently as an associate professor in the Department of Biological Sciences. While at Kent State, he developed a new microscopic method to measure cell volume, and that prompted him to turn to the physiology of cell volume regulation. He is also interested in the biological role and control of macromolecular crowding and continues to work on microscopic techniques and their applications.

Affiliations and expertise

Kent State University

Irena Levitan

Irena Levitan, PhD, is a Professor of Medicine and Bioengineering at the University of Illinois at Chicago. She received her PhD in Biophysics and Neurobiology at the Hebrew University of Jerusalem in 1994 and completed postdoctoral training at the Medical College of Pennsylvania and Institute for Medicine and Engineering at the University of Pennsylvania. Her research focuses on the biophysical properties of endothelial membranes and sub-membrane cytoskeleton. Specifically, the studies of her group, which combine computational and experimental biophysical approaches, provided the first comprehensive structural insights into cholesterol regulation of K+ channels. In 2012, she was named a Guyton Distinguished Lecturer “for her quantitative and biophysical work on cholesterol modulation of ion channels and how this can affect integrated organ function”. She and her group also discovered a paradoxical relationship between fluidity/deformability of the membrane and cell stiffness. In 2018, she was elected a Fellow of AIMBE for “outstanding contributions to our understanding of lipid-ion channel interactions, cellular biomechanics and vascular dysfunction under dyslipidemia”.

Affiliations and expertise

Professor of Medicine and Adjunct Professor of Pharmacology and Bioengineering, University of Illinois, Chicago, USA