Breaking flow cytometric paradigms

Grant Howes of Accuri Cytometers explains why a multi-faceted single instrument is most efficient for researchers.

Today's life science researchers utilize technologies from many disciplines. Purchasing then mastering varied instruments and technologies devours time and monetary resources. Often a technique may be needed in only a single experimental phase leading to infrequently used instruments cluttering valuable bench space. The ideal solution is an instrument, which can perform a myriad of multi-parametric applications. A flow cytometer fits that description but, to-date, flow cytometry has not enjoyed the mainstream acceptance primarily due to the cost, size and complexity of existing instruments. As with so many technologies, manufacturers have added a seemingly endless series of bells and whistles to mature platforms over time, rendering them increasingly unwieldy. Conventional flow cytometers are expensive, large, high-maintenance instruments that require extensive training to use correctly.

The Accuri C6 Flow Cytometer System breaks the flow cytometry paradigm. With a footprint, weight and price similar to a real-time qPCR system or microplate reader, the C6 puts an essential cell analysis tool into the hands of more biologists than ever before. The instrument offers all the performance features of larger, complex flow cytometers and does not require expert operators or dedicated staff, making flow cytometry equally available to both novices and experts.

Researchers have used flow cytometry for an expanding application set for decades. This quantitative, analytical technology is often used to study cellular phenomena that might also be investigated by microscopy, microplate reading, qPCR or Western blots. Beyond classical immunophenotyping, such experiments include cell cycle analysis, apoptosis, cell proliferation and viability as well as assessment of transfection efficiency and molecular-level changes in phosphorylation and methylation states, cellular and molecular profiling and rare cell screening. Current choices in kits, buffers, antibody-fluorochrome conjugates and protocols make designing and implementing multicolor assays easier than ever. The proliferation of multiplex bead-based assays has allowed flow cytometers to replace time-consuming, commonplace assays such as ELISAs, and opened the door for flow cytometry to be used much more widely in the fields of gene and protein microarray analysis

Due to the wide variations in fluorescence and light scatter signals produced by different types of cells and particles, flow cytometrists often spend significant time and valuable sample, optimizing amplifier gains and/or PMT voltage settings. These settings cannot be changed post-analysis if signals were improperly amplified, resulting in the irretrievable loss of data.

The Accuri C6 Flow Cytometer is equipped with pre-optimized detectors, calibrated to operate within their linear range, so it can be used to analyze a wide variety of samples, ranging from dim, barely-fluorescent, micron-sized platelets through large, >30 micron, highly-fluorescent cell lines. The combination of high-resolution digital-signal processing (24-bit DSP), resulting in an expansive dynamic range on all detectors, and the CFlow software capability to 'zoom' in on very small areas of data display make the C6 easy to use for both novices and experienced users. The C6 allows the separation of data acquisition and data analysis by obviating the need for setting voltages. This advance allows novices to be able to collect high-quality data with minimal direction or supervision and to set gating strategies and fluorescence compensation values during, or after, data collection. Data collected on a C6 is always retained and can be re-analyzed, at any time, if gating or compensation errors are discovered, or in light of new research findings.

The Accuri C6 Flow Cytometer offers unique advantages over more traditional methods by enabling multi-parametric, individual cell analysis. With a linear dynamic range over six decades, the C6 quantitatively captures the entire scope of biological variations in a single run without the need for data acquisition optimization or tuning. The C6 can also quantitatively measure the concentration of cells or particles in samples and correlate this with the specific known volumes being sampled. Multi-parametric flow cytometric analysis can now replace other less accurate techniques and be accomplished as needs dictate. By designing a 2-laser, 6-detector, affordable flow cytometer, Accuri has successfully addressed the mainstream cell biology applications with a single platform and freed up valuable resources.

For more information please visit www.accuricytometers.com.

Grant Howes is Marketing Director at Accuri Cytometers, Inc. An industry veteran, Grant has worked in cell analysis for over 30 years of cell analysis experience and has close to 25 years of flow cytometry experience in both the research and clinical marketplaces.