A New Alternative for High-Throughput Screening

The Accuri C6 Flow Cytometer® System combined with the IntelliCyt™ HyperCyt® Autosampler runs a 384-well plate in ten minutes and provides an affordable flow-based platform for the University of Michigan’s High-Throughput Screening Core.

“Twenty 384-well plates are currently run daily on the Accuri C6 HyperCyt combination. We could run up to 40 plates in a standard 8 hour workday, over 12,000 compounds”
-Martha Larsen, Director, HTS, siRNA, HCS Laboratory

Cell versus Well
Fluorescent plate readers provide aggregate assay results for each well in a multi-well plate. These aggregate results reflect the average characteristics of the population and could mask relevant subpopulations that may be of particular importance to the subject under investigation.

In contrast, flow cytometers analyze cells or particles on a singular basis providing cell-by-cell statistics that allow differentiation between cell subpopulations. This makes it possible to derive information about the physical and chemical nature of each individual cell or particle.

The University of Michigan's High-Throughput Screening Core Facility
The High-Throughput Screening (HTS) core facility, a central component of the University of Michigan's Center for Chemical Genomics (CCG), www.lsi.umich.edu/facultyresearch/centers/chemicalgenomics, is designed to assist academic researchers in carrying out high-throughput screens of chemical libraries to identify new tools for biological research. The HTS core facility acts as a regional resource and assists in assay development and primary screening, dose response confirmation and structure activity analyses.

Approximately 100 researchers use the HTS core facility.

When the HTS core facility was initially established, they focused on providing "wellular" data using PHERAstar and FlexStation® 3 plate readers. Needs changed in 2007 when a principal investigator's research required cellular data analysis. Resources were pooled and the HTS core facility purchased an Accuri C6 Flow Cytometer System and IntelliCyt HyperCyt Autosampler for high-throughput cellular and multiplex bead analysis.

To Date, Nearly 200,000 Compounds Have Been Screened
"I did not want another complicated piece of instrumentation to learn and maintain," explained Martha Larsen, Director, HTS, siRNA, HCS Laboratory. "As it turned out the Accuri C6 is not a complex instrument, is easy-to-use and it is also easy to train new users on. The C6 and HyperCyt combination is an excellent addition to the core facility and gives us the capability to provide cellular and bead-based data. Together the two instruments only take up about 4 feet of bench space. The C6 SIP is easily attached to the HyperCyt so that we can use the instruments together or use the C6 as a stand-alone flow cytometer for smaller sample numbers."

Finding a high-throughput system was crucial to the HTS core facility, not only for speed, but also to reduce the sample volume needed for analysis; assay costs increase as sample volumes increase. The HTS core facility uses a 5 µL/well sample size. The small amounts of carryover observed initially have been minimized and Z-factors on assay plates have run consistently at 0.7-0.8 indicating high-quality screens.

Twenty 384-well plates are run daily on the Accuri C6 HyperCyt combination. (A 384-well plate run takes 10 minutes; a 96-well plate 2.5 minutes.) To date, three targets have been screened. The only drawback is that plates must be changed manually. The HTS core facility plans to address this in the future by integrating the system with a robotic arm.

Simple connections allow you to switch between plate mode and normal tube sampling in minutes. This ability to connect and disconnect the two instruments is crucial to the HTS core facility. A little over half of their walk-up users only have a handful of samples and prefer to use the C6 for cellular analysis in a non-high-throughput mode.

The Accuri C6 and HyperCyt combination has expanded the capabilities of the HTS core facility to provide cellular and multiplex bead analysis. When not needed for screening, the C6 acts as a stand-alone flow cytometer for smaller sample numbers or assay development activities.

Software integration is complete and available off-the-shelf. Accuri Cytometers and IntelliCyt Corporation worked together to allow the C6 and HyperCyt to communicate and to run on a single computer. The C6 software, CFlow® for the HyperCyt System, collects the data then autoexports it in FCS format to the HyperCyt software, HyperView®, for data analysis.

Minimal Downtime
A core facility loses money when there is instrument downtime or when hours are spent trouble shooting. "Accuri's Technical Support group has been very quick to resolve the few issues that have popped up," Martha Larsen commented. "The group works with us over the phone or, since we are only located about 15 minutes away from one another, sends a technical representative, if necessary."

The robustness, size and reliability of the C6 allow Accuri to offer a depot repair policy. Within 48 hours of a decision by the Accuri Technical Support team that the instrument must be returned for service, Accuri sends a loaner C6 and picks up the original instrument. Repairs are made and the original and loan instruments are swapped back.

Accuri C6 Flow Cytometer System and IntelliCyt HyperCyt Autosampler
A high-performance flow cytometer simplified, the Accuri C6 Flow Cytometer System features two lasers, two light scatter and four fluorescence channels. Designed specifically with the life scientist in mind and priced comparable to a fluorescent plate reader, the C6 has a small footprint and no special set-up requirements. With built-in capabilities to measure cell concentration, the C6 is a perfect tool for cell counts and can accommodate large assay runs (proliferation, viability, cell cycle, etc.) with the optional, automated CSampler™.

For laboratories that want the walk-away convenience that automation provides but do not need high throughput, the recently released CSampler is an ideal solution. CSampler is less than half the price of other flow cytometry automation systems and processes a 96-well plate in approximately 90 minutes.

HyperCyt, on the other hand, was developed specifically to accelerate the rate of sample delivery to standard flow cytometers. This unique patented technology addresses the need for speed and efficiency. By speeding up the delivery of samples to flow cytometers and streamlining data analysis, HyperCyt makes high-throughput flow cytometry a practical reality for applications in drug discovery, systems biology and life sciences research.

The HyperCyt is compatible with the Accuri C6 and most other flow cytometers.

Accuri, C6 Flow Cytometer are registered trademarks of Accuri Cytometers, Inc. HyperCyt is a registered trademark of IntelliCyt Corporation. FlexStation is a registered trademark of Molecular Devices.