Single Cell Biology

Currently, the redox state of single, living cells is accessible only by fluorescence microscopy. Upon oxidation or reduction of key amino acids associated with fluorescent redox proteins, conformation changes cause differences in emission wavelength. Thus, the relative amounts of oxidized and reduced protein can be used to gauge the redox environment of a cell. Similarly, the redox state can be studied if the emitting protein is specific to a certain metabolite species, as has previously been shown (Cambronne et. al., 2016, Science) for cytosolic NAD+. Unfortunately, these experiments cannot be performed in the absence of light. It has been shown that the act of shining light on a cell can have a deleterious effect on metabolism. For instance, cytochrome C absorbs blue light and degrades, decreasing mitochondrial respiration with time. We are developing nanoelectrode probes to study how wavelength and intensity of light affects cell metabolism at the single cell level. Furthermore, we are fabricating metabolite-specific nanoelectrodes. This research can be used across a vast range of cell lines to study variations in metabolism across different areas of human disease. 

4 Beads Gif

Hyperspectral Imaging

Within only a few seconds, we can image several fluorescently-tagged species. Pictured here for proof-of-concept are four polystyrene beads each labeled with a different fluorophore.

Hyperspectral Imaging

Our lab is interested in making highly quantitative measurements inside single, living cells. This requires development of specialized microscopy tools, specifically those that have white light laser capabilities without the expense of white light lasers.

Nanoelectrode

This SEM micrograph of a nanoelectrode shows the sharp, pointy tip after pulling with a laser-puller. Sharp electrodes are able to penetrate a cell and make sensitive measurements within a cell.

Cells

While imaging cells is commonplace in most cell biology experiments, the effect of wavelength and intensity of light on cellular metabolism is still an open question.

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© 2019 by Laboratory of Jeffrey E. Dick, All Rights Reserved

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