The BD FACSCanto™ is often hailed as the new generation of flow cytometry. One striking difference between the FACSCanto™ and older generation machines such as the FACSCalibur™ is that this system operates on a PC-based Windows XP® platform using BD FACSDiva™ software. These tools assist in the ease and efficiency of using FACSCanto™ as they enable automated features. Techniques used to prepare cell samples for FACSCalibur™ are the same as those used for FACSCanto™; the only difference is the need to learn how to use FACSDiva™ software, which assumes you have a working knowledge of basic Microsoft Windows® operating systems.
The BD FACSCanto™ combines a patented design for enhanced signal retention on six fluorescent and two scatter parameters, digital electronics for processing up to 10,000 events per second, and a novel sample injection tube supporting carryover of less than 0.1%. Additional benefits include the best fluorescence sensitivity for detection of dim events, automated compensation algorithms with optional off-line compensation, streamlined sample loading, and automated housekeeping routines.
The BD FACSCanto™ system is built with blue (488nm, air-cooled, 20 mW solid state) and red (633nm, 17 mW HeNe) excitation sources. Laser beams are routed using fiber optics; the lasers beam shaping prisms which are projected onto separate spots on the flow cell. The emission signals are transmitted from the flow cell to the detector arrays, which control the sensitivity of this system. Digital electronics have improved the performance and made the instrument much easier to use. The electronics eliminate dead time and the need for interbeam time-delay calibration. The digital electronics facilitate compensation, with no limits to inter- and intra- beam compensation, allowing post acquisition compensation.
Perhaps, one of the most remarkable aspects of this system is the fluidics system, which has been designed to save time and improve performance. Increased volumes of fluids can be stored, thereby eliminating the need to interrupt data acquisition to refill the saline tank. The system performs an automated flush between samples to assist in reducing sample-to-sample contamination. Additionally there is no more need to worry about losing your sample when a tube is inserted onto a needle. The sample is more likely to become diluted rather than lost.
Our lab currently uses the FACSCanto™ for various applications, ranging from immunophenotyping assays to multicolor analysis. Since using the FACSCanto™, we have been able to acquire more data from each experiment, thanks to the increased number of parameters available for acquisition and analysis. This and the increased speed have proven to be the biggest advantages when using this machine. The FACSCanto™ is, therefore, an excellent investment for any core facility doing routine flow cytometry work.
Joudy-Ann Dinnall
Pre-Doctoral Fellow
The Children’s Hospital of Philadelphia
Division of Rheumatology