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Flow Cytometry Technique Enables High-Speed Cell Sorting

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A study led by global medical technology company BD (Becton, Dickinson, and Co.), in collaboration with the European Molecular Biology Laboratory (EMBL), has demonstrated an innovation in flow cytometry that adds fluorescence imaging and image-based fluorescence to sort individual cells at exceptionally high speed based on the visual details of each cell, as opposed to on the type or quantity of biomarkers that are present.

Traditional methods of cell sorting through flow cytometry operate through the identification and quantification of certain biomarkers (for example, proteins) on or within a cell. The method described in the study, called BD CellView Image Technology, can capture multiple images of individual cells flowing through the system at a speed of 15,000 cells per second, and sort them based on detailed microscopic image analysis — something that was previously impossible.

The technology has applications in immunology, cell biology, and genomics research, and it could enable new cell-based therapeutic discovery.

Image-enabled cell sorting identifies and isolates cells of interest out of a complex pool of cells at speeds up to 15,000 cells per second. Courtesy of Tobias Wuestefeld/BlueClay Studios.


Image-enabled cell sorting identifies and isolates cells of interest out of a complex pool of cells at speeds up to 15,000 cells per second. Courtesy of Tobias Wuestefeld/BlueClay Studios.
By adding imaging to the traditional biomarker identification and quantification, the technology not only identifies if and how much of a biomarker is present in a cell, but also its location and how it is distributed within the cell. By imaging the distribution of biomarkers with this technology, researchers obtained detailed information about cells that was previously invisible in traditional flow cytometry experiments. That information enables investigation of complex biological questions, such as how cells grow, function, and interact, and/or to study the exact location of viruses or proteins within a cell, all at a highly accelerated pace.

“This innovation has overcome the typical compromise between speed and precision of sorting individual cells,” said Tom Polen, chairman, CEO, and president of BD. “This breakthrough essentially equates to a researcher looking into a microscope, identifying specific characteristics of a cell of interest, and, based on what they see, sorting each individual cell for further analysis — all at a rate of nearly 1 million cells every minute.”

Using the technology, it is possible to analyze more than 1000 times the amount of data of which traditional flow cytometry methods are capable of analyzing, Polen said. It is also possible to sort cells at a rate of 15,000 per second based on their images.

In the study, researchers used BD CellView Image Technology to study regulators of the NF-κB (nuclear factor kappa light chain enhancer of activated B cells) pathway, a protein complex that plays an important role in cellular immunity and stress response. The EMBL team measured the activity in this pathway by tracking the location of RelA, a protein that moves from the cytoplasm into the nucleus of the cell upon activation.

Using BD CellView Image Technology, the screen allowed them to identify several novel regulators of this important cellular pathway in a matter of hours, instead of days as would be the case using conventional approaches.  

The new technology fills a long-standing gap in biomedical research by enabling scientists to more rapidly view and isolate cells with specific, observable traits of interest, which can accelerate discovery research and unlock potential therapies or cures for disease in a broad range of fields such as virology and oncology.

“This technology represents the culmination of more than a decade’s worth of work from a highly multidisciplinary team of optical, mechanical, electrical, biomedical, software engineers, and scientists that aimed to provide researchers a differentiated and flexible capability for analyzing single cells,” said Eric Diebold, worldwide vice president of R&D for BD Biosciences and co-corresponding author of the paper. “We have just scratched the surface of the types of science that will be enabled with this new high-throughput image-based cell sorting technology, and we look forward to how BD and the scientific community at large will leverage it to advance both basic research and the development of therapeutics.”  

The research was published in Science (www.doi.org/10.1126/science.abj3013).

BioPhotonics
Mar/Apr 2022
GLOSSARY
flow cytometry
A method of measuring the characteristics of microscopic particles, usually cells, as they flow in a fluid stream through a beam of light. Particles may be stained with fluorescent dye and the fluorescence detected via laser illumination.
cell
1. A single unit in a device for changing radiant energy to electrical energy or for controlling current flow in a circuit. 2. A single unit in a device whose resistance varies with radiant energy. 3. A single unit of a battery, primary or secondary, for converting chemical energy into electrical energy. 4. A simple unit of storage in a computer. 5. A limited region of space. 6. Part of a lens barrel holding one or more lenses.
Research & Technologyflow cytometryimagingBiophotonicsCellcell countingcell sortingBDEuropean Molecular Biology LaboratoryEMBLEuropecollaborationsciencecell imagingBioScan

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