E. Paluch – Biophysical Methods in Cell Biology (2015)

Автор: E. Paluch
Название книги: Biophysical Methods in Cell Biology (2015)
Формат: PDF
Жанр: Биологические науки
Страницы: 513
Качество: Изначально компьютерное, E-book

This new volume of Methods in Cell Biology looks at methods for analyzing of biophysical methods in cell biology. Chapters cover such topics as AFM, traction force microscopy, digital holographic microscopy, single molecule imaging, video force microscopy and 3D multicolor super-resolution screening
Covers sections on model systems and functional studies, imaging-based approaches and emerging studies
Chapters are written by experts in the field
Cutting-edge material

Biophysical approaches have recently been taking an increasingly prominent place
in the cell biology toolbox. Historically, physics, and specifically mechanics, has
been tightly intertwined with biology since the early days of cell and developmental
biology. Throughout the twentieth century, embryologists have been using glass capillaries,
needles, and deformable plates to probe and perturb the mechanical properties
of cells. With the expansion of optical microscopy techniques in cell biology,
these approaches have been complemented with optical perturbation and quantitative
imaging. In this volume, we cover a variety of biophysical methods allowing
for the quantification and perturbation of the organization, dynamics, and mechanics
of structures at the subcellular, cellular, and tissue scales. Far from being exhaustive,
the chapters presented here intend to cover various aspects of the cell biophysics
toolbox and to highlight recent developments in the field. Many of the methods
rely on custom-made software that is either available online or can be obtained
from the authors upon request. This highlights the growing need for flexible and
adaptable computer software in cell biology. Computational approaches go hand
in hand with an increasing use of modeling in cell biology. While physical models
are often too context-specific to lend themselves to a method chapter on modeling
itself, many of the methods presented here, particularly those dealing with the measurement
of cellular mechanics, rely on a model for the extraction of physical quantities
from quantitative data. Such cross talk between quantitative experiments and
modeling is becoming increasingly central to modern cell biology.
The first nine chapters present different aspects of how high-end optical techniques
can be used for quantitative cell and developmental biology. These include
methods for single-molecule imaging of dynamic processes in live cells (Chapters
1e3), and quantitative methods for characterizing the organization, morphology,
and dynamics of intracellular structures, such as cytoskeletal or mitochondrial networks
(Chapters 4e6). The following two chapters describe the application of novel
superresolution imaging techniques for high-throughput screening (Chapter 7) and
imaging of cellular structures in vivo, in the Drosophila embryo (Chapter 8). Finally,
Chapter 9 presents the implementation and various applications of digital holographic
microscopy for the measurement of the cellular refractive index. The following 10
chapters present methods for the measurement of cellular mechanical properties.
Chapter 10 describes the development and implementation of FRET sensors for the
measurement of tension at the molecular level. Chapters 11e13 focus on measurements
of cellular physical properties using deformable plates (Chapter 11) and
atomic force microscopy (Chapters 12 and 13). Chapter 14 presents a pipette-based
method for probing cellecell adhesion. Chapters 15e17 describe the implementation
of different traction force microscopy methods for mapping forces exerted by
cells on substrates. Chapters 18 and 19 present methods for the quantification of
cellular strains and stresses from quantitative imaging of cell aggregates. Finally,
Chapter 20 presents a method using oil droplets as probes to measure cell-generated
stresses. Besides methods for measuring dynamics and mechanics, recent years have
seen an increasing use of biophysical techniques to perturb and manipulate cell
behavior. Chapters 21 and 22 describe techniques for controlled perturbation of
plasma membrane mechanics and integrity. Chapters 23 and 24 present examples
of how microfabrication can be used to study specific cell biology questions. Note
that a recent Methods in Cell Biology volume extensively describes various applications
of micropatterning in cell biology (Methods in Cell Biology, volumes 119, 120,
and 121 edited by Matthieu Piel and Manuel The´ry). Finally, Chapters 25 and 26
describe easy to implement methods for the quantification of global cell dynamics