Development of chemical probes for live cell Raman imaging
EMSL Project ID
60899
Abstract
Biological molecular phenotyping requires characterization tools that can record multiple biological processes and molecular transformations simultaneously in real time. Currently, fluorescence based chemical probes are used to measure enzyme activity, and metal ion concentration. Mass spectrometry and RNA based fluorescent probes are used to measure metabolites. While these probe types have advanced our understanding of biological systems significantly, the broad spectral features associated with florescent dyes limit the number of biological processes and solutes that can measured simultaneously, and 2) mass spectrometry sampling is nominally destructive for soft matter, 3) Moreover, the lack of straightforward and high-throughput methods for visualization for metabolites of interest in live cells (e.g., RNA based probes do not allow measurement of metabolite concentration inside the cell because of the large size of the probe) represents a current bioimaging bottleneck. Thus, there is a need to develop novel methods and materials that allow measurements of enzymatic activity, metal ions and metabolites simultaneously in a non-destructive, high-throughput fashion in live cells. Recent advances in chemistry related to Raman active tags (e.g., polyynes) improved their sensitivity and have allowed use in sub-micromolar concentrations of Raman probes for imaging. We will take advantage of this new chemistry and develop novel chemical probes to measure enzyme activity, metal ions and metabolites using stimulated Raman spectroscopy. The probes will be built with Raman active chemical groups such as alkyne including polyynes or nitrile groups which are optimal in this application because they appear at the biologically silent region of the Raman spectra. With this current project, we will expand EMSL’s chemical probe-based imaging capability to include chemically selective, multiplexed live cell imaging.
Project Details
Start Date
2023-10-01
End Date
N/A
Status
Active
Released Data Link
Team
Principal Investigator
Team Members