LIVE-PAINT: New Imaging Technique Revealing Workings of Living Cells
A new imaging technique that could pave the way to a better understanding of many diseases, a technique that reveals the inner workings of living cells in stunning detail has been developed by Edinburgh scientists.
A flexible and powerful way of tracking individual proteins inside living cells, without disrupting their activity is provided by the new super-resolution imaging technique, called the LIVE-PAINT.
This technique can reveal the behavior of proteins involved in disease processes and those essential to health and can be a game-changing advance in leading to new insights into diseases.
The existing super-resolution techniques involve a fusion of a large fluorescent molecule that can interfere with the protein of interest, require complex instruments, or cannot be used inside living cells, thus they are limited.
The LIVE-PAINT technique was developed by the team at the University of Edinburgh, and by tagging a protein-of-interest with a very small protein molecule, known as a peptide, they tested it in live yeast cells.
At the University of Edinburgh’s School of Biological Sciences, Professor Lynne Regan, the senior author of the study explains, “The chances of a tag affecting the protein’s natural function is minimized in this technique, as the LIVE-PAINT offers a way to label proteins for super-resolution imaging with a very small tag.”
Then, the peptide tag temporarily binds to a peptide-binding protein, which is fused to a fluorescent protein. The necessary blinking to construct super-resolution images is generated by this reversible binding, as it drives the repeated association of the fluorescent protein to the protein-of-interest.
In diseases that are characterized by the harmful build-up of protein aggregates inside cells, the approach could offer new insights, such as Alzheimer’s, ALS, and motor neuron disease.
As existing super-resolution methods often behave differently when directly fused to a large fluorescent protein, using them to study the proteins involved in aggregation diseases is difficult.
LIVE-PAINT could be used to study many other important processes in living cells without disrupting their activity and it requires the fusion of a small peptide.
A detailed study of a group of medically important proteins, which are the target of 70% of current drugs known as transmembrane proteins can be made possible through this approach, for the first time. Embedded in the membrane surrounding cells, transmembrane proteins have vital roles, such as signaling between nerve cells, and when they are disrupted, it often leads to disease.
Another technique, which uses short DNA strands to reversibly associate a fluorescent molecule to a target molecule, the super-resolution technique called DNA-PAINT is where the LIVE-PAINT approach takes inspiration from.
In DNA-PAINT technique, the short strands of DNA can only be used in dead cells that have been broken up or on the outer surface of a cell, as they are synthesized chemically. While in LIVE-PAINT, using the cell’s own protein production machinery, all the components are made inside the cell.
At the University of Edinburgh’s School of Chemistry, Dr. Matthew Horrocks, the senior author of the study said, ” LIVE-PAINT will be accessible to many groups, not just specialists, given its straightforward implementation, and we particularly pleased that the versatility of LIVE-PAINT.”
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LIVE-PAINT: New Imaging Technique Revealing Workings of Living Cells
