WOODBURY, NY — CompuCyte this week held a scientific symposium and demonstration at Cold Spring Harbor Laboratory's Genome Research Center here as part of its effort to support the iCys quantitative imaging cytometer installed at CSHL's core flow cytometry facility last year.
CompuCyte, based in Cambridge, Mass., has organized symposia and user group meetings in the past for its 'i' Series imaging cytometers, but for a much broader audience. The CSHL meeting was notable because it was the first time the company held such an intensive meeting for a group of users from a single facility, Elena Holden, CompuCyte's president and CEO, told CBA News.
The CSHL symposium demonstrated the wide array of application areas for which CompuCyte's newest instruments are suited, but also revealed one of the biggest challenges for the relatively small company: With so many potential markets to play in, how to focus on the most important ones?
The answer to this is still a work in progress, and the company will be able to better resolve these areas as it obtains feedback from multiple users at core facilities such as CSHL. But for now, according to Holden, CompuCyte continues to position its instruments as an alternative or complement to traditional flow cytometers.
Pam Moody, manager of the flow cytometry facility at CSHL, said that one research lab at her institute purchased the iCys early last year to support a specific research project. Since then, the instrument has found its way into the core cytometry facility, and Moody has slowly learned many of its features, even if other facility users have not.
"Flow cytometry in general is of course one area where we compete very well, but even that is somewhat complementary."
She told CBA News that she hoped the symposium and accompanying demonstrations would help CSHL researchers better understand the versatility of the iCys instrument, which combines many of the features of traditional flow cytometry with those of an automated microscopy platform.
"People have been asking 'What does it do?'" Moody said. "But not as many people have tried it out as we would have liked. And it's not a very intuitive technology [for someone] coming from a flow background. Even though it combines flow and imaging microscopy, there are a lot of things in it that you can only learn by watching someone or doing it yourself a few times.
"Basically, when you learn this instrument, you're supposed to walk first before you run, and it kind of went from getting the instrument to trying to run with it," she added.
Moody said that after the symposium, there are now about a dozen researchers at CSHL that have expressed interest in using the iCys for their research projects, and she expects many more to come forward in the next few months. She added that there is the potential for well over 100 users, which is approximately how many people use the lab's Becton Dickinson LSR II flow cytometer.
"There were about six people at the seminar from this institution alone that had never set foot in the cytometry facility, so the interest is there," Moody said. Examples of projects CSHL researchers are now interested in include cell-cycle analysis, fixed tissue analysis, stem cell studies, fluorescence resonance energy transfer assays, and intra-nuclear labeling assays, among others, she added.
Moody's comments underscore one of the biggest challenges facing CompuCyte: determining the most promising markets for its imaging cytometers.
CompuCyte's Holden said the company is still learning about applications from customers, but in the meantime believes that it can play in the same space as traditional flow cytometers.
But competing for budget dollars with flow cytometers is not easy, either, as that technology is well established, and has been a staple of research labs for about three decades.
"The versatility and flexibility are key strengths of the instruments, but marketing is one of the biggest and most exciting challenges," Holden said. "It does well in the areas where you require a high degree of flexibility, such as in core facilities. Flow cytometry in general is of course one area where we compete very well, but even that is somewhat complementary."
CompuCyte's challenges are similar to those of Amnis, a Seattle-based biotech also marketing a quantitative imaging cytometer, but there's one major difference: Amnis' instrument images non-adherent cells in flow, while CompuCyte's instruments are designed to provide quantitative measurements and images of cells or tissue in well plates, culture plates, or slides.
Not surprisingly, Amnis, which recently pocketed more than $11 million from a private equity financing round, is also positioning its instrument as an alternative or complement to traditional flow cytometers (see related story, this issue).
Speakers at the CSHL symposium discussed the way they use the iCys and iCyte platforms. One was William Telford, director of the core flow cytometry facility at the National Cancer Institute. The NCI facility was one of the earliest adopters of CompuCyte's instrument, having bought one in 1999 when it was called the LSC II. The facility upgraded that instrument last year to the next-generation iCys.
Telford's presentation covered proper fluorochrome selection for CompuCyte's imaging cytometers, which use three laser lines — 488 nm, 633 nm (or an optional 594 nm), and 405 nm — and therefore can only excite dyes with similar excitation spectra.
Furthermore, he discussed how users should take into account the photobleaching time of dyes when using laser-scanning cytometers like CompuCyte's.
"In flow, we don't often think about photobleaching, because the laser dwell time is very short," Telford said. "With laser-scanning cytometry, like confocal microscopy, the dwell time is longer and can cause photobleaching, and rescanning the sample is also typical."
Telford also said that quantum dots are becoming very popular for use with flow cytometers, and similarly are more often being paired with LSC instruments.
One of the biggest projects involving the iCys at NCI is a project with the National Institute of Standards and Technology called the "biomarkers initiative," in which researchers are using the instrument to investigate Her-2-neu expression in breast cancer tumor lines through immunohistochemistry.
Zbigniew Darzynkiewicz of the Brander Cancer Institute of New York Medical College discussed how his group is using the iCys measure DNA fragmentation during apoptosis or in response to anti-tumor drugs or radiation.
Darzynkiewicz said that there were several reasons his group uses the iCys over traditional flow cytometry, such as the ability to measure time-resolved events in individual cells; the ability to re-analyze data from specific cells; and the ability to obtain information on cellular and tissue architecture.
A pair of CompuCyte's pharmaceutical customers discussed how the company's instruments can further drug-discovery research such as toxicity testing and biomarker studies. Catherine Hu, a research scientist at GlaxoSmithKline, talked about how her group has used a CompuCyte platform to quantitatively assess mitochondrial function and apoptosis in rabbit parietal cells in 96-well plates to investigate gastric toxicity.
Lastly, Amy Shen from Pfizer presented work that her research group had done using an iCyte to detect both fluorescent and chromogenic biomarkers for liver toxicity using whole tissue slices and tissue microarrays.
Holden told CBA News that CompuCyte's imaging cytometers are well-suited for all of these application areas, and are all representative of the type of work that will pass through a core cytometry facility.
"Our strategy is to continue to find out where [the instruments] are working the best, and to pursue those areas," Holden said. "We know that the instrument is not best-suited for something like conducting multiple translocation assays, although we have shown that it can be used for that type of work."
— Ben Butkus ([email protected])