The Accuri C6 can measure volume, and therefore allows calculation of cell concentrations within CFlow Plus software statistics tables without the addition of counting beads. This eliminates sources of variability associated with dual platform counting and the expense of counting beads.

Click here for more information.

Many fluorescent protein (FP) variants can be analysed using the Accuri C6 Flow Cytometer System. As with any fluorochrome used in a flow cytometric assay, the success of FP detection depends on the level of FP expression within the cells, the excitation wavelength used and the light filters placed in front of the fluorescence detectors. This technical note provides guidelines for the C6 user performing FP analysis.

Background

The successful cloning and expression of the green fluorescent protein (GFP), derived from the jellyfish Aequoria victoria (Chalfie et al., 1994 and Tsien, 1998), ushered in a new era for fluorescence microscopic studies of living cells and their functional processes. Since the initial use of GFP in gene expression studies, mutagenesis has been used to produce a number of variants (e.g., eGFP, eCFP, eYFP) with varying emission maxima across the blue to yellow wavelengths. In 1999, the gene for a red fluorescent protein (RFP) from the sea anemone Discosoma striata was cloned (Matz et al. 1999). Subsequent mutagenesis of this FP has also yielded a wide range of derivatives which have extended the useful spectral emission range into the red region (Baird et al., 2000). Many of these GFP and RFP derivatives have proven useful not only for fluorescence microscopy but flow cytometry as well. The combined use of flow cytometry and FP technologies has been successfully employed in almost every field of biology, including physiology, cell biology, molecular biology, immunology, stem cell research, and microbiology.