Other Publications (1)
Articles by Pravat Dhakal in JoVE
Ratiometric Calcium Imaging of Individual Neurons in Behaving Caenorhabditis Elegans Bhavya Ravi*1, Layla M. Nassar*1,2, Richard J. Kopchock III*2, Pravat Dhakal*2, Michael Scheetz2, Kevin M. Collins1,2 1Neuroscience Program, University of Miami School of Medicine, 2Department of Biology, University of Miami This protocol describes the use of genetically encoded Ca2+ reporters to record changes in neural activity in behaving Caenorhabditis elegans worms.
Other articles by Pravat Dhakal on PubMed
Bioimaging with Macromolecular Probes Incorporating Multiple BODIPY Fluorophores Bioconjugate Chemistry. | Pubmed ID: 28430413 Seven macromolecular constructs incorporating multiple borondipyrromethene (BODIPY) fluorophores along a common poly(methacrylate) backbone with decyl and oligo(ethylene glycol) side chains were synthesized. The hydrophilic oligo(ethylene glycol) components impose solubility in aqueous environment on the overall assembly. The hydrophobic decyl chains effectively insulate the fluorophores from each other to prevent detrimental interchromophoric interactions and preserve their photophysical properties. As a result, the brightness of these multicomponent assemblies is approximately three times greater than that of a model BODIPY monomer. Such a high brightness level is maintained even after injection of the macromolecular probes in living nematodes, allowing their visualization with a significant improvement in signal-to-noise ratio, relative to the model monomer, and no cytotoxic or behavioral effects. The covalent scaffold of these macromolecular constructs also permits their subsequent conjugation to secondary antibodies. The covalent attachment of polymer and biomolecule does not hinder the targeting ability of the latter and the resulting bioconjugates can be exploited to stain the tubulin structure of model cells to enable their visualization with optimal signal-to-noise ratios. These results demonstrate that this particular structural design for the incorporation of multiple chromophores within the same covalent construct is a viable one to preserve the photophysical properties of the emissive species and enable the assembly of bioimaging probes with enhanced brightness.