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Mar 15, 2013 (Vol. 33, No. 6)

Democratizing Flow Cytometry

  • Life Technologies’ offerings include the Attune® acoustic focusing flow cytometer, which provides sample throughput at rates up to 1,000 µL/minute. Applications that can be carried out on this benchtop system include apoptosis analysis, cell-cycle analysis, phosphoprotein detection, immunophenotyping, rare event detection, intracellular marker detection, basic phenotyping (up to six colors), cell proliferation detection, green and red fluorescent protein detection, live/dead cell discrimination, and phagocytosis detection.

    Sony Biotechnology’s vp for marketing and distributor operations, Lew McAllan, sees a number of novel applications coming to the fore with the new compact flow cytometry instruments. These fluorescent protein analysis, particularly simultaneous multifluorescent protein analysis and sorting, and stem cell analysis and sorting. Among Sony’s current product is the new SH800, a personal cell sorter that comes with up to four lasers and six-color detection.

    The SH800 incorporates a microfluidic chip that replaces the conventional flow cell and nozzle design, with the goal being to ensure a clean system.

    “The SH800 fully automates the setup for cell sorting,” explained McAllan. “In addition, a small size is achieved by taking advantage of the integrated laser technology and compact mechanical design technology cultivated in the Sony Blu-ray optical techniques.”

    He noted that Sony also is developing a spectral analyzer, that by virtue of a 32-channel PMT and deconvolution algorithms in software, will allow simultaneous analysis of overlapping fluorochromes, providing at least 14-color analysis for two standard lasers.

  • Smaller Can Be Better

    Click Image To Enlarge +
    BD Biosciences’ recent advances in personal flow cytometry are enabling high-accuracy, high-purity single-cell isolation for analysis of gene expression and next-generation DNA sequencing.

    J. Clark Mason, Ph.D., senior director of global marketing for flow cytometry at BD Biosciences, emphasized that BD always focuses on developing a complete system when it is updating its flow cytometry technology. He said that lasers packing the same power have been made smaller allowing for more compact instruments with a higher performance/footprint. These instruments can “punch beyond their weight,” he said.

    In addition, there has been a revolution in the power of the fluorescent dyes that can be used. Last August BD acquired the Sirigen Group, including its polymer dye products. According to Dr. Mason these are 10–20X brighter than conventional fluorescent dyes and have much reduced complication of spectral overlap.

    “Such dyes add to the power of the new technology platforms, providing the capacity for increased multiplexing and permitting identification of dimly staining populations that were previously challenging to resolve,” he said, adding that the development of more sophisticated software permits researchers to carry out in-depth analysis on the smaller, benchtop platforms.

    Dr. Mason said that recent advances in personal flow cytometry are contributing to the revolution in the genomics arena where the technology is enabling high-accuracy, high-purity single-cell isolation for analysis of single-cell gene expression and next-generation DNA sequencing. According to Dr. Mason, BD recently launched a benchtop cell sorter—FACSJazz™—that makes commonly used cell-sorting applications accessible to researchers with limited flow cytometry experience.

  • One Technology Led to Another

    Miltenyi Biotec’s involvement in flow cytometry dates back some 20 years to the company’s origin specializing in magnetic cell separation (MACS®), explained Shane Oram, Ph.D., global marketing manager, flow cytometry. The technology was used to evaluate the quality of separation that had been achieved via the magnetic technique.

    Dissatisfaction with the instruments available at the time led Miltenyi to develop its own line of flow cytometry instruments.

    “Today’s benchtop instruments do virtually everything for the user who simply needs to push a few buttons and walk away,” he said.

    With its power to analyze hundreds of thousands of cells in minutes, flow cytometry instruments can replace microscopes for many applications, added Martin Büscher, Ph.D., head of biophysics at Miltenyi. The systems are now used in stem cell research, organelle analysis, circulating tumor cell identification, water monitoring, the checking of lipid content of biofuels, and many other areas, he pointed out.

    Dr. Oram described Miltenyi Biotec’s latest flow cytometer, the MACSQuant® VYB, as a three-laser, eight-color benchtop analyzer that has a pipetting robot on the front end and also offers magnetic cell separation to pre-enrich samples when looking for rare events.

    According to Dr. Büscher, Miltenyi Biotec, as well as some other companies, now offer calibration particles that can be used by researchers to ensure consistent and comparable results from day-to-day and from lab-to-lab.

    Suzanne Hancock, project manager in the R&D group for research and applied markets at GE Healthcare Life Sciences, said that the company had recently conducted an extensive customer survey of a wide range of scientists to assess their flow cytometry needs. The results indicated that the scientists were highly interested in simplification of the technology and in hassle-free systems that would allow them to do every-day tasks that might normally require the use of three to four different instruments and would pull all these tasks together on one platform.

    The scientists also were looking for increased efficiency from both research acceleration and manufacturing productivity standpoints. They want systems with greater accessibility that could be used in the individual lab to provide immediate answers to typical questions such as the health of the cells, number of cells, and phenotypes, and thus allow the researchers to move on to the next experiment, rather than having to wait for an answer to come back from the core lab.

    In addition, the researchers have a need for systems that can efficiently handle small volumes of material as they often did not have the luxury of having significant sample quantities. Systems that would permit them to implement their own applications as well as providing preset standard applications were also on the researchers’ list of needs.

    In response, Hancock explained that GE Healthcare developed a new benchtop platform, the Cytell™ Image Cytometer, which was introduced to the marketplace last year. This instrument is actually not a flow cytometer, but performs many of the same functions and provides an actual image of the cells. Applications include analysis of physical characteristics, cell counting and viability, apoptosis analysis, assessment of cell health, and phenotypic analysis of fluorescent proteins and cell surface markers.

    The instrument is intended to supplement standard flow cytometers. Numerous applications are preconfigured on the system and simply require the pushing of a button to implement, according to Hancock.

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