To develop the advanced flow cytometry process that is at the crux of Medusa and may one day be part of an advanced astronaut health-monitoring system, NASA sought the help of private industry. The Agency issued a solicitation through the Small Business Innovation Research (SBIR) program at Ames to find a partner for the project. Specifically, NASA needed a partner that could produce a high-speed flow cytometry process to continuously monitor cells for anomalies and bacteria growth. The system would ideally be able to image cells with high-fluorescence sensitivity and would be tolerant of wide variations in sample concentration and other characteristics by having a wide depth of field, allowing cells to be kept in focus regardless of where they happened to be in the flow stream.

The submergence vehicle Alvin as it is about to dive to the bottom of the ocean. Released from the mothership, the Research Vessel Atlantis (operated by Woods Hole Oceanographic Institution, of Woods Hole, Massachusetts) with the diver standing on top.
Flow cytometry is a powerful technique, but it has several limitations that hinder its use for NASA projects. Most commercial flow cytometers cannot image cells like a microscope. Instead, they measure only the total amount of fluorescence emitted by each cell. Because they do not have the ability to determine where in the cell the signal is coming from, their applications in cell biology are mainly limited to measuring the total quantities of specific molecules in or on the cell. A few flow cytometry systems can produce images of cells in flow using transmitted light, but these systems lack the sensitivity necessary to image faint fluorescence.

Screen shot of Amnis Corporation’s IDEAS data analysis software showing the detection of cells in division.
Flow cytometers also generally require that cells flow through the center of a tightly focused laser beam, which can make them vulnerable to misalignment. Designs that are less sensitive to misalignment tend to sacrifice fluorescence sensitivity.

Many flow cytometry systems are also just too big and impractical for NASA's purposes, especially since they incorporate pressurized fluid vessels that employ gravity and high pressure to drive the sample through the system.

Amnis Corporation, a Seattle-based biotechnology company, developed a technology called ImageStream for producing sensitive fluorescence images of cells in flow, and happened to be seeking ways to get whole cells into focus in order to increase the usefulness of its systems for research applications. The company had several ideas for how to achieve an extended depth of field, all of which required a level of funding that was just not in its budget. When Amnis heard about the SBIR solicitation, however, the realization came that it could be the perfect opportunity to reap the funding necessary to develop extended depth-of-field technology. The company responded to the SBIR solicitation and proposed to evaluate several methods of extending the depth of field for its ImageStream system, pick the best method, and implement it as an upgrade to its commercial products. This would allow users to view whole cells at the same time, rather than just one section of each cell.

Through Phase I and II SBIR contracts, Ames provided Amnis the funding the company needed to develop this extended functionality. For NASA, the resulting high-speed image flow cytometry process made its way into Medusa and has the potential to benefit space flight health monitoring. On the commercial end, Amnis has implemented the process into its flagship product, ImageStream.