Confocal microscopy is an analytical technique routinely used in the Mirkin group to characterize the activity of our many biologically-active nanoconjugates, including cellular uptake, intracellular analyte detection, and subcellular compartmentalization. However, the microscope currently in use, a Zeiss 510 purchased in early 2008, is unsuited for live cell imaging. This microscope suffers from extremely slow acquisition times, which, for live cell confocal microscopy means long exposure times to high-powered, damaging lasers. Currently, high levels of “phototoxicity” from each exposure kills live cells after just a few images, making it impossible to perform multiple-image analyses such as time-lapse imaging or 3D reconstruction. Furthermore, when analyzing live cells that continuously move and change, slow acquisition time yields low quality, often inaccurate images. In recent years the Mirkin group has significantly increased the number of projects involving biological systems, including multiple DARPA funded projects that involve analyzing the ways in which our nanoconjugates interact with cells over time. Currently, our use of a microscope that is not designed for live cell analysis is a significant limiting factor to the progress of these projects. Since 2008 significant advancements in confocal microscope technology have been made, resulting in microscopes that take images over three orders of magnitude faster than our currently-used Zeiss 510, with greater clarity, and operate at laser powers that do not harm cells. Specifically, the opportunity to use a Zeiss Cell Observer SD spinning disk confocal microscope would allow for live cell imaging at a level of clarity and complexity far beyond our current abilities. Furthermore, the motorized, programmable stage included with the system will allow us to analyze multiple samples in parallel. When used in the planned time-lapse imaging over periods of 48 hours or more, this feature will drastically decrease data acquisition time. This state-of-the-art instrument will additionally allow the training of DOD students and postdocs in advanced fluorescence microscopy techniques and image analysis.
|Effective start/end date||8/28/14 → 8/27/15|
- U.S. Air Force Asian Office of Aerospace Research and Development (FA2386-14-1-3004)
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