Flow cytometry companies have been making news lately, with new products, agreements and an acquisition. Advances in lasers, detectors and reagents have allowed for more complex and sensitive analysis. Applications of flow cytometry include apoptosis, cell-cycle phosphorylation status, surface markers, protein expression and DNA-content determination.

In 2009, the market for research flow cytometers is valued at more than $750 million. Driving market growth are new systems, including those with higher-throughput capabilities and lower-cost instrumentation, and new applications, such as those utilizing multicolor flow cytometry and those utilizing 96-well plates.

Beckman Coulter is one of the two dominant providers of research and clinical flow cytometers. Its research flow cytometry product line includes the MoFlo XDP Cell Sorter, the high-speed CyAn, the compact Quanta and the newly released, modular Gallios. Recently, the company entered into two agreements to increase the high-throughput capabilities for the market. In early August, the company signed an agreement to marry Intellicyt’s Hypercyt autosampler, which can process a 96-well plate in two-and-a half minutes and a 384-well plate in 10 minutes, with the CyAn high-speed digital research cytometer, which can analyze 80,000 events per second (eps). This agreement enables the CyAn, which previously retrieved samples from tubes, to process 96- and 384-well plates. Dr. Richard Kendall, director of Global Marketing, Flow Cytometry Business Center at Beckman Coulter, described the advantages of high-throughput flow cytometry to IBO. “Whereas traditional high-throughput screening (HTS) has relied on single end-point biochemistries, HTS by flow cytometry allows for much more sophisticated multiparametric measurements that cover a variety of cell functions. This combination will provide opportunities for major academic institutions and for pharma and biopharma companies to use flow cytometry as a mainstream high-speed screening tool.” Ninety-six well-plate reading in flow cytometry is currently used for secondary screens and biomarker analysis, but its use as a primary screening tool is relatively new. Beckman Coulter also now holds exclusive distribution rights for the HyperCyt outside of North America, and the co-exclusive distribution rights for the HyperCyt system for use with its CyAn flow cytometer in North America. The company’s second high-throughput agreement was for the MoFlo system. Under a new agreement with Beckman, Propel Labs will upgrade existing MoFlo systems to the MoFlo XDP model, a higher-throughput version of the MoFlo with an analysis rate of 100,000 eps and a sort rate of 70,000 eps.

Beckman Coulter also introduced a new flow cytometer, the Gallios, in March. The standard two-laser Gallios is available in three configurations and can be outfitted with an optional violet laser, and 6-10 fluorescence detectors. The system’s increased sensitivity is designed to handle complex experiments undertaken with newer reagents. “The Gallios brings novel opportunities to resolve dimly fluorescent signals that challenge the extremes of high-speed cytometry,” explained Dr. Kendall. “As cytometry moves towards the temporal detection of brightly labeled surface markers and dimly staining intracellular signals, the challenge has become to resolve both simultaneously. The Gallios meets these challenges in newly emerging applications like cell signaling and intracellular cytokine staining.” These complex applications are also highly amenable to multicolor cytometry, both of which are growing the market.

The introduction of the Gallios further increases Beckman’s offerings for multicolor flow cytometry. “The ability to simultaneously detect multiple biological signals gives researchers the opportunity to understand cellular functions in ever increasing granularity. Subfractions of subfractions of individual cellular populations can be simultaneously studied for both intracellular and extracellular markers, or functions that may change in response to a variety of agonists or stimuli.” Phenotyping of multiple antigen-specific T-cells is one particular application that benefits from the addition of lasers and detectors to cytometry systems, because a majority of T-cell subsets can only be identified through the simultaneous discrimination of four or more molecules’ expression patterns. Dr. Kendall cited other benefits of multicolor flow cytometry as well. “Other advantages include the efficiencies of being able to use the same cells labeled with many markers to reduce the unnecessarily repetitive use of monoclonal antibodies. It also permits the preservation of scarce cells and can reduce the number of pipetting steps required to perform an analysis.”

Millipore entered the flow cytometry instrument market earlier this year by acquiring Guava Technologies, an established provider of low-cost flow cytometers (see IBO 2/15/09). Guava’s product line consists of the easyCyte, easyCyte Mini and the PCA-96 systems. Unlike other flow cytometers, Guava’s systems use microcapillary flow cells in place of sheath fluid to carry the sample through a laser beam where fluorescent dyes are excited. According to Guava, the sheathless design allows its systems to operate with less sample and generate less waste. With the acquisition, Millipore’s sizeable resources can expand the distribution and development of Guava’s systems. “Guava Technologies has long been trying to broaden the use of flow cytometry within individual labs by providing instruments with specific assays and software analysis tools,” said John Sweeney, vice president of Millipore’s Life Science Strategic Business Unit. “However, they did not have the resources to drive a broader adoption into the wider research market and could only focus around specific research areas.” Mr. Sweeny also made clear that Millipore’s increased R&D funding for flow cytometry increases the company’s capabilities in cellular analysis. This includes “cell cycle and cell growth, immunology, cell function and activation, cell differentiation, cell signalling, GFP detection, and toxicology,” according to Mr. Sweeny.

Millipore’s newest flow cytometry instrument, released in July, is the easyCyte 8HT. According to the company, the system is available for nearly half the cost of traditional two-laser and four- to six-detector systems. In contrast to Guava’s other systems, the easyCyte 8HT is equipped with a blue and a red laser (versus a blue or a green laser), and can analyze up to six targets and eight parameters (versus up to three targets and five parameters). The system is also highly automated. In addition to its ability to obtain samples from tubes, it comes with a 96-well plate reader. The software allows for a plate to be read and displayed as a heat map, rather than providing only a single-well reading. As a result, different experiments in multiple wells run on the same plate and trends in a single experiment run on the plate based on multiple parameters can be identified.

As part of its acquisition of Guava, Millipore stated that one of the company’s goals would be to integrate its reagent kits with Guava’s instruments. One specific example of this is the FlowCellect kit for stem cell analysis, which was introduced in response to customers requests for additional methods to monitor the phenotype of stem cells. “Millipore was the first company to provide such assay solutions. While these have recently been copied by market leaders within the flow cytometry market space, Millipore is also able to serve researchers by providing adjacent products such as those related to the culture of stem cells, which differentiates us from competitors,” said Mr. Sweeny. Millipore also plans to further expand flow cytometry’s presence in the biotech and pharmaceutical bioprocess markets, where the Guava instruments are currently used to optimize and monitor cell culture conditions. “The constant pressure within this industry is rapid, in-time manufacturing data. We will be exploring the use of this technology to provide quick data and information pertaining to manufacturing so that the correct decisions can be made before extra costs are incurred, thereby benefiting in time and cost savings,” explained Mr. Sweeny.

Accuri Cytometers is one of the newer manufacturers of flow cytometry systems. The company released its first product, the C6 flow cytometer, in April 2008. Perhaps the most distinctive feature of the C6 is its $40,000 price tag. Jennifer Baird, Accuri president and CEO, stated that the largest stumbling block that the company faced in introducing the system was convincing researchers that the system’s price did not indicate inferior performance. Unlike Guava’s flow cytometers, the C6 operates with a sheath fluid. It comes equipped with blue and red lasers and four fluorescence detectors. “The C6 can be used in the manner that’s similar to conventional flow cytometers, but we have innovations in fluidics, electronics and optics that have miniaturized the platform. So we have eliminated the need to set voltages and gains on our system, which much simplifies application work that needs to be done…That is enabled further by the fact that we have industry leading dynamic range that makes it simpler for people to use,” explained Ms. Barid. The company also rececntly added automation to its system. In May, it released the CSampler, which adds a 48- and 96-well plate-reading capability.

The C6’s lower price has also allowed Accuri to introduce the system to markets that have not traditionally used flow cytometers. “We are taking flow cytometry to different places, such as agriculture, food-safety monitoring, research on growth patterns of algae for biofuel production and community colleges. Basically, a number of places that flow cytometers are not found today,” said Ms. Baird. However, as she emphasized, the C6’s innovations and price does not mean that it only competes with low-cost flow cytometers. “We compete with two-laser, four-color flow cytometers that are out there. They can be everything from Becton Dickinson to Beckman Coulter’s market-leading machines. Essentially, most of our competitor’s machines are well over $100,000, and our price point is a fraction of that.” She also described how the C6 is capable of competing in the multicolor cytometry market, stating that the company’s research shows that 90% of multicolor cytometry is still done with four colors.

Chart: Total Research Flow Cytometry Market Value

Total ($ Millions)

2008 $780

2009 $760

2010 $850