With the new method, the microscope is bridged on the spot with single cell ohms.
Scientists can now select individual cells from the growing population on the surface of laboratory utensils to study their molecular contents. A new tool developed by researchers at the University of Toronto will allow in-depth study of stem cells and other rare cell types for the development of therapy.
The method is the first to link the cell microscope to ohmic platforms, the physical parameters of cells that are visible to the naked eye, such as the appearance of surface marks or cell contacts, to connect them to their molecular composition.
“We allow the user to take beautiful images of a fluorescent microscope to learn everything that can be learned about cells growing locally, and then link that information to the cell genome, transcriptome, or proteome,” said Aaron Wheeler, a professor of chemistry. Biomedical Engineering at Donell Center for Cell Biomolecular Research, who supervised the work.
The platform is described in an article published in the journal today (November 11, 2020) Nature communications,
Called DISCO, for digital microfluidic isolation of single cells for ohms, the method allows researchers to select single cells in their cells և to analyze their contents DNA: և Cell DNA (genome), genes RNA transcripts և protein molecules (proteome).
The growth of single-cell analysis over the past five years has allowed researchers to measure tens of thousands of molecules in each cell, transforming their ability to study tissues and organs at the granular level. But these approaches use important information about the physical properties of the cells և the local environment, as the cells must be suspended and separated before they can be analyzed.
“Now there’s a revolution in cell dissection,” said Wheeler, a Canadian research fellow in microfluidic bioanalysis. “But I met people who were frustrated by the fact that they were not able to capture phenotypic information about the cell in its in situ environment.”
“And I thought we might be able to find a way to select certain cells from that population and analyze them,” he said.
DISCO consists of a microscope equipped with high-frequency lasers and a microfluidic chip for collecting cellular material. The microscope allows the user to take a detailed picture of the target cell before the laser beams on it. The energy from the laser causes a small bubble to explode next to the cell, tearing its membrane, firing its contents into a microchip chip from which it is taken for molecular sequence.
“Our platform focuses on the metadata that you lose when you make a single cell suspension, things like the position of the cell, what were its morphological properties, what were its neighbors. “These are all things we can do before completing a single cell sequence,” said Erika Scott, a lab graduate researcher who co-led the lab with two graduate students, Julian Lamanna and Harrison Edwards.
“According to our information, this is the only platform that can take cells in culture to do such things,” he said.
Demonstrating groundbreaking experiments, the researchers demonstrated DISCO’s ability to faithfully link omics data to individual մարդու mouse brain cancer cells developed side by side.
But the findings have also focused on the extent to which interactions between cells can affect their molecular states. The expression of the mouse’s 5,000 genes, about one-fifth of the genome, was altered in individual cells of mice surrounded by human cells instead of their relatives.
The findings could have repercussions for many laboratories trying to better understand healthy, diseased human tissue, such as tumors, by growing them in mice so that they can be studied throughout the body. If the expression of the gene affects the human vaccine in the same way, those changes could have consequences for the development of treatment, Wheeler said.
Fortunately, DISCO may soon be able to offer a window into cells in their natural environment as researchers work to adapt it to tissue fragment analysis. Their ultimate goal is to use DISCO to study rare cell types, such as stem cells, whose regenerative potential is largely regulated by their immediate environment to help advance new therapies.
Reference. Julian Lamanna, Erika Y. Scotty, Harrison S. Edwards, M .; Dean Chamberlain, Michael D. Մ. Dryden, Iaaki Peng, Barbara Meyer, Adam Lee, Calvin Chan, Alexandros A. Sklavounos, Austin Heffernan, Farhana Abbas, Charis Lam, Maxwell E. Olson, Jason Moffat and Aaron R. Wheeler, 11 November 2020 Nature communications,
DOI: 10.1038 / s41467-020-19394-5: