Cells
are dynamic
environments that use carefully
regulated mechanisms to maintain function and health. One example of
this is
the vesicle-mediated transport of lipids (shown to the right). Each
bright spot
shows a single vesicle as it transports lipids through the cell. Each
step of
this process; internalization, transport in the vesicle, and enzymatic
degradation of the lipids, is controlled by chemical reactions and
mechanical forces within the cell. Understanding these dynamic
processes
requires a method that will provide both spatial and temporal
information-the
ability to watch each step as it occurs. To obtain this information we
use
fluorescence microscopy to directly probe intracellular dynamics. The
Payne
Lab is interested in two related questions; what are the rates and
mechanisms of these
intracellular processes and how can we better image each event.
Direct imaging
reveals the subcellular location,
concentration, and reaction rate of the molecules of interest. These
parameters can be measured to determine a complete intracellular
reaction mechanism.
Specific systems of interest include post-translational modification of
proteins, transcytosis across the blood-brain barrier, and delivery of
cargo to the lysosome for degradation. These systems pose a number of
biological and physical questions including the mechanism of
intracellular
transport, kinetics of vesicle fusion, influence of the local
environment on a
chemical reaction, and the conversion of chemical energy into
mechanical
motion.
New methods
for live cell imaging:
Fluorescence microscopy and nanomaterial delivery.
The Payne Lab is
developing new optical techniques for
live cell imaging and new methods for delivering novel fluorescent
probes to
cells. These methods will be used to probe intracellular reactions on
the
molecular level and to enable new research directions using
quantitative
cellular imaging. Optical methods of interest include nanometer-level
imaging,
spectroscopic single-particle tracking, and multiphoton total internal
reflection. Intracellular delivery of novel fluorescent probes and
other nanomaterials will
borrow
methods developed for gene delivery to introduce synthetic materials
into cells in a
controlled manner.