Payne Lab

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[[Payne Lab | <font face="trebuchet ms" style="color:#ffffff"> '''Home''' </font>]] &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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='''Imaging Reaction Dynamics in Living Cells'''=
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='''Imaging Chemical Reactions in Living Cells'''=
[[Image:cell_rxns_paynelab.gif|right|390 px]]
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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.
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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 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 the Payne Lab uses fluorescence microscopy to directly probe intracellular dynamics.  
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==='''Intracellular degradation of extracellular cargo'''===
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==='''Imaging chemical reactions within a cell.'''===
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Cells control the chemical reactions responsible for the utilization of nutrients, replication of viruses, and regulation of receptors through the localization of substrates and enzymes within distinct vesicles that are actively transported through the cell. We are especially interested in the reactions that result from the interaction of substrate-containing vesicles with enzyme-containing vesicles. We are using two-color single particle tracking to address this question in standard cell lines and in a cellular model of the blood-brain barrier.
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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.
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==='''Nanoparticle-cell interactions'''===
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Nanoparticles have important biomedical applications ranging from the treatment of human disease with gene therapy to understanding basic cellular functions with fluorescent probes. For these applications to be fully realized it is necessary to understand how nanoparticles interact with cells. The Payne Lab is especially interested in the cellular binding and internalization of nanoparticles in the presences of extracellular proteins. A combination of advanced microscopy and spectroscopy techniques are being used to understand these fundamental interactions.
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==='''New methods for live cell imaging: Fluorescence microscopy and nanomaterial delivery.'''===
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==='''New methods for live cell imaging'''===
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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. The intracellular delivery of novel fluorescent probes will borrow methods developed for gene delivery to introduce fluorescent probes and other nanomaterials into cells in a controlled manner.
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While recent developments in fluorescence microscopy make it possible to image many of the dynamic events that are essential to cellular function, new methods are necessary to observe the dynamics of single molecules inside living cells. Imaging within live cells is difficult as the emission from fluorescent probes competes with the autofluorescence of the cell. The Payne Lab is developing new optical techniques for quantitative cellular imaging. Optical methods of interest include nanometer-level imaging, spectroscopic single-particle tracking, and multiphoton total internal reflection microscopy.

Revision as of 11:09, 15 May 2012

Research         People         Publications         Funding         News         Seminars         Positions Available         Outreach         Contact        


Contents

Imaging Chemical Reactions in Living Cells

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 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 the Payne Lab uses fluorescence microscopy to directly probe intracellular dynamics.

Intracellular degradation of extracellular cargo

Cells control the chemical reactions responsible for the utilization of nutrients, replication of viruses, and regulation of receptors through the localization of substrates and enzymes within distinct vesicles that are actively transported through the cell. We are especially interested in the reactions that result from the interaction of substrate-containing vesicles with enzyme-containing vesicles. We are using two-color single particle tracking to address this question in standard cell lines and in a cellular model of the blood-brain barrier.

Nanoparticle-cell interactions

Nanoparticles have important biomedical applications ranging from the treatment of human disease with gene therapy to understanding basic cellular functions with fluorescent probes. For these applications to be fully realized it is necessary to understand how nanoparticles interact with cells. The Payne Lab is especially interested in the cellular binding and internalization of nanoparticles in the presences of extracellular proteins. A combination of advanced microscopy and spectroscopy techniques are being used to understand these fundamental interactions.

New methods for live cell imaging

While recent developments in fluorescence microscopy make it possible to image many of the dynamic events that are essential to cellular function, new methods are necessary to observe the dynamics of single molecules inside living cells. Imaging within live cells is difficult as the emission from fluorescent probes competes with the autofluorescence of the cell. The Payne Lab is developing new optical techniques for quantitative cellular imaging. Optical methods of interest include nanometer-level imaging, spectroscopic single-particle tracking, and multiphoton total internal reflection microscopy.

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