Problem: Flow cytometry is a powerful technology that allows researchers and clinicians to perform complex cellular analysis quickly and efficiently by analyzing several parameters simultaneously. The amount of information obtained from a single sample can be further expanded by using multiple fluorescent reagents. The number and combinations of fluorescent reagents that can be used by a flow cytometer depend on the types of lasers, filters and detectors with which each instrument is equipped. The more lasers and detectors an instrument has, the more simultaneous detection of colors it is capable of. However, by increasing the number of colors (reagents), researchers are also incorporating additional levels of complexity.

Solution: Multicolor flow cytometry enables more data to be gathered about a sample in a shorter amount of time, giving researchers not only enhanced efficiency and high-quality data, but also more data from less sample than what would be required for serial analysis. There are several factors researchers should consider before developing a multicolor flow cytometry assay with more colors in a single tube.

Instrument type: The type of instrument that the researcher has access to, and its availability, will define how many and what colors can be used in the experiment.

Experimental need: Some simple assays require only two to four colors, so researchers would not need to consider more colors.

Availability of reagents: Once a researcher knows which colors his or her instrument supports and which antibody markers are required, he or she will need to identify reagents conjugated to an appropriate fluorochrome.

Expertise in multicolor panel design:Many researchers do not have the experience or technical understanding to put the ideal or most appropriate color combinations together that will maximize their chance at success. This can lengthen the time it takes to get results.

Advanced flow cytometry tools are being developed that will allow researchers to use multiple colors more effectively. BD Biosciences offers a comprehensive set of multicolor solutions, including cutting-edge flow cytometry instruments and software, high-quality reagents and knowledgeable technical applications support.

BD FACS™ (fluorescence-activated cell sorter) brand flow cytometry instruments support a growing number of multicolor applications and are ideal choices for multicolor flow cytometry. BD flow cytometers, including the BD FACSCanto™, BD FACSAria™ II and BD™ LSR II systems, provide the reliability and versatility needed for use today and in the future. In fact, the BD LSR II benchtop flow cytometer processes 70,000 events per second, measuring up to 18 fluorescent colors.

BD FACS DIVA 6.1 software includes a CS&T (cytometer setup and tracking) feature that, in combination with the BD CS&T Beads, automatically sets up and tracks the day-to-day performance of the cytometers and adjusts automatically to ensure reduced variation and reproducible results in your multicolor flow cytometry assays over time.

For more information, go to www.bdbiosciences.com/colors