Seattle-based Amnis has sold two of its ImageStream 100 imaging flow cytometers to academic core cytometry labs at the University of Pittsburgh Cancer Institute and the University of Minnesota for use in basic research, Cell-Based Assay News has learned. The sale brings to six the total number of disclosed customers of the platform since it was officially launched in December
In addition, Amnis is poised this month to announce the signing of an $860,000, 18-month Phase II Small Business Innovation Research contract from the National Institute of Environmental and Health Sciences to use its ImageStream in order to develop high-throughput fluorescence in situ hybridization assays of mice sperm, David Basiji, Amnis' chief technology officer, told Cell-Based Assay News.
The two announcements underscore the wide variety of potential applications for the relatively young instrument platform, a mix between a flow cytometer and a fluorescence microscope.
However, the instrument sales are a step toward the company's recently stated objective of placing the ImageStream in basic research laboratories and core flow cytometry labs, the two most suitable venues for the ImageStream's capabilities, Jack Ball, Amnis' president and CEO, told Cell-Based Assay News in December (see CBA News, 11/23/2004).
The aforementioned universities join the Fred Hutchinson Cancer Center, the National Institute of Allergy and Infectious Disease, biotech firm Cell Therapeutics, and the Coriell Institute for Medical Research as customers that Amnis has announced thus far.
"The nature of the instrument and its cost ends up qualifying it most strongly for core labs, although there are a number of individual labs that are well-motivated and well-financed that have purchased them, as well," Basiji added last week. "But I think it's safe to say that most ImageStream 100's will end up in core labs."
According to Basiji, the University of Minnesota platform has just been installed, but Amnis has not yet conducted user training. Meanwhile, the UPCI has been using the ImageStream for a little over a month.
"We had training in February, and it was installed in March," Albert Donnenberg, a professor of medicine at the University of Pittsburgh and director of UPCI's flow cytometry facility, told Cell-Based Assay News. "We actually were not one of the beta-testers. David Basiji came out and gave a talk late last year, and we were blown away by the ImageStream's capabilities.
"The cancer center is leasing the instrument right now, but we have put a grant in to the NIH for funding," Donnenberg added. "We couldn't wait to get it."
Donnenberg said that although UPCI is the customer, the core facility in which the ImageStream resides is open to all UPitt researchers. As Donnenberg sees it, Amnis offers two main advantages over any existing instrument.
"First, you can associate fluorescence with cellular features. This allows you to do translocation-type assays," he said. "For instance, we have researchers that are working on NFkB … that are using the instrument."
The second feature is something Amnis calls virtual sorting, where, using conventional flow analysis, researchers can sort cells based on specific features within the images. An example of where this would be important, Donnenberg said, is in cancer cell differentiation work.
"There's something called the stem-cell hypothesis for cancer, which in a nutshell is that tumors have heterogeneity with respect to differentiation, and at the bottom of it all is essentially a small stem cell," he said. "We're trying to construct a differentiation tree based on this theory, and the technology is very useful for these studies. It allows you to associate markers with cell morphology." Donnenberg also said that the ability to distinguish between subsets of cells is useful in applications involving immune cells such as B-cells and T-cells.
Many of these types of applications can also be performed on high-content imaging systems such as Cellomics' ArrayScan, GE Healthcare's IN Cell Analyzer, Evotec's Opera, and Molecular Devices ImageXpress or Discovery-1.
Basiji nevertheless maintains that Amnis views its main competition to be flow cytometry, not high-content screening companies.
"That's the user base that understands what we do the quickest, and seems to be the fastest to take up the technology," Basiji said. "We often get asked about how we compare to high-content screening systems. And it's really apples to oranges, because we're not in the business of chugging through hundreds of thousands of samples a day, with a couple hundred cells per sample.
"It's much more of an in-depth tool for a fewer number of samples," he added. "So it's for discovery biology, it's great for [toxicity] studies, and if you've got a couple of dozen promising leads — it's great for that. But we're not going to be cranking through plates with this thing."
While the ImageStream may not be high-content per se, it certainly provides higher content than a flow cytometer.
"I do put us in a different class than a flow cytometer," Basiji said. "We get hundreds of times more information from a cell sample. But we do get everything that a flow cytometer gets, and we do handle cells in flow, and we can run pretty much any standard protocol that anybody has developed for flow cytometry over the last 30 years — any of the dyes, any of the cell-probing protocols. So it's a natural comparison."
"The ability to look at a large number of cells with this — up to 50,000 per run — is a huge benefit," Donnenberg said. "That number is not a lot for flow cytometry, per se, but it is a huge number for imaging."
Amnis' pending NIEHS contract is the second grant Amnis has received this year related to FISH applications on the ImageStream.
In February, the company said it has won an 18-month, $600,000 Phase II grant from the National Aeronautics and Space Administration to develop an extended depth-of-field version of ImageStream, which may eventually be used to study the effects of microgravity and cosmic radiation exposure on the cells of astronauts — a timely grant considering the US government's recent outlining of a plan for a mission to Mars.
"Right now we get 4 to 5 microns, at most, for depth of field in the cell," Basiji said. "[Currently, people analyze those signals manually on a microscope. By having the extended depth of field version of our platform, we'd be able to keep the entire cell and nucleus in focus the entire time."
In the case of the NIEHS contract, which has yet to be officially announced by the company, Amnis is expected to develop a commercial platform for sperm analysis, [including] protocols and reagent kits.
"It's mainly for aneuploidy detection, where the NIEHS is trying to characterize various mutagens and teratagens, and quantitate how frequently they give rise to genetic mutations in the germ line," Basiji said.
Basiji said that researchers typically expose mice to different chemicals at different doses, and then do "thousands and thousands of FISH experiments to try and detect aneuploidy in their sperm.
"Again, it's incredibly labor-intensive," Basiji said. "I don't know off the top of my head how long it takes them to characterize one of these compounds, but it's long. They'd like a system that can do this with high throughput, so they can crank through a number of mutagens and teratagens, and really characterize their toxicity."
Even though the grant is specifically for murine sperm analysis, Basiji said that Amnis thinks it might be able to exploit the same capability for fertility testing in human sperm, citing recent evidence that suggests aneuploidy in sperm — in addition to ova — may be correlated to a decrease in human fertility with age.
However, Rifaat Salem, medical director of the Pacific Reproductive Center in Torrance, Calif., told Cell-Based Assay News that such an application may not be that promising, because it is still by and far aneuploidy in the ova that correlates strongest with infertility.
"Considering age in relationship to aneuploidy, at age 35 in women, it's only about 30 or 40 percent aneuploidy, but when women reach age 40, this becomes over 60 percent," Salem said. "There are a few papers regarding sperm, but women are born with all of their eggs, but men can make a brand new batch of sperm every three months, so men are not as affected. You can get viable sperm from men at age 40, 50, 60, and so on."
Still, clinical researchers conduct FISH experiments on samples obtained from ova, so there may be some potential down the road for the ImageStream to make some impact in infertility testing.
"FISH in general is still an area that has tremendous clinical potential and commercial potential, but it's also an area that's going to require more work before it's ready for the market," Basiji said. "That's why we're getting grants in that area. In terms of commercial success [of the ImageStream], it's clearly in the general research market and the drug discovery process."
Scott Keefer, a senior application scientist at Cellomics who is a certified clinical lab specialist in cytogenetics, told Cell-Based Assay News that, in theory, Amnis' technology could be useful in high-throughput FISH analysis.
"The classic way to do this is to gain a sample of blood or whatever it may be, and then manually score whether it's a positive or negative score," Keefer said. "The throughput of this is still pretty low … and accuracy is the key. You also don't want to introduce any bias. So by automating this, you take out the human error, which is a big thing. So there could definitely be advantages to this."
However, Keefer added, "Not only Cellomics, but a lot of the other competitors just haven't gone after the diagnostics market, because there's not a whole lot of money there. Other high-content screening platforms could probably do this sort of thing as well, but they've been focusing primarily on the drug-discovery process because that's where the money is."
Furthermore, Keefer added, companies such as Applied Imaging have been doing automated microscopic scoring of FISH for many years.
Amnis' platform, however, would likely offer higher throughput than existing platforms because of its flow capabilities.
"You could, in a single path through the instrument, detect all the FISH spots from the cell, and … turn this into a really high-throughput technique to look for relatively rare genetic problems," Basiji said. "We routinely run 10,000 cells through our system in a couple of minutes. Compared to a microscope looking at 50 cells on a slide, this could be a lot more effective as a diagnostic test."