Previously, the images of distant galaxies were only seen as blurry spots. Researchers at Washington University in St. Louis have developed a new method for showing the contents of a single cell.
The researchers were led by Srikanth Singamaneni, a professor of mechanical engineering and materials science in the school. A single cell can be seen and measured in about 30 minutes.
In collaboration with researchers in the Washington University School of Medicine and other universities, they found that the FluoroDOT test is versatile, low-cost and can be used in a variety of laboratories. Current methods are limited in sensitivity and can take up to 24 hours to process.
The plasmonic-fluor is 16,000 times brighter than conventional fluorescent labels and has a signal-to-noise ratio of nearly
"Plasmonic-fluors are composed of metal nanoparticles that serve as antenna to pull in the light and enhance the fluorescent emission of the molecule," Singamaneni said.
This ultrabright emission of plasmonic-fluor allows the user to see extremely small quantities of secretedprotein, which they are unable to do in existing assays, and measure the high-resolution signals digitally using the number of particles, or dot pattern, per cluster, or spot, using Special equipment is not required. The plasmonic-fluor work was published in 2020.
The office of technology management at Washington University licensed the plasmonic fluor technology to Auragent Bioscience.
"We are able to simultaneously image a cell along with the spatial distribution of the proteins secreted around it, using a simple fluorescence microscope, and we continue to work on it as a principal scientist." Different cell types showed interesting secretion patterns. Two types of proteins can be visualized from individual cells. When multiple cells are exposed to the same stimuli, we can distinguish between the cells that are secreting two genes at the same time from the cells that are only secreting one.
The team used cells from both humans and mice to test the technology.
The co-director of the Division of Infectious Diseases in the School of Medicine, and one of the co-author's, has used the FluoroDOT test.
"When the immune cells are exposed to Mycobacterium Tuberculosis, they respond by secreting important immune genes, called cytokines," he said. Some cells respond differently to infections. We were able to see which cells were secreting and which direction they were in with the help of the Fluoro DOT. The older technology did not allow this.
More information: Anushree Seth et al, High-resolution imaging of protein secretion at the single-cell level using plasmon-enhanced FluoroDOT assay, Cell Reports Methods (2022). DOI: 10.1016/j.crmeth.2022.100267 Journal information: Cell Reports Methods , Nature Biomedical Engineering
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