1Multicycle AV for DNA/Cell Cycle Analysis

FCS Express provides a complete DNA analysis solution based upon the widely used Multicycle AV™ DNA analysis package written by Dr. Peter Rabinovitch at the University of Washington, and distributed by Phoenix Flow Systems.

Multicycle is included with FCS Express and reports if more than one cycle is detected in your .fcs data files, and/or if a background correction would improve the fit. Both of these features are available in FCS Express using the Multicycle tool to ensure accurate analysis.

2Proliferation Analysis

Assays to assess the proliferative activity of cells are commonly used for monitoring the health and growth rate of cell populations.  In this type of assay, a cell tracking dye, such as CFSE, is incorporated into the cells.  With each round of cell division, the relative fluorescent intensity of the dye decreases by half.

As an initially homogeneous population of cells divides, you will get a mixture of  cells containing a half, a quarter, an eighth, a sixteenth, etc., of the initial population. Different conditions often cause a population of cells to change their proliferation characteristics, and understanding these effects is important in many different fields.

FCS Express uses a sophisticated linear least-squares fitting process to extract the correct information from your data and separate the populations.

3Kinetics Analysis

Kinetics experiments are particularly useful for examining changes in DNA content, antibody binding, and calcium flux over time.  For example, calcium ions play an important role in cell signaling. The intracellular concentration of calcium ions often show transient changes in response to external stimuli. Several fluorescent dyes can be used to measure bound Ca++ by flow cytometry, including Indo-1 (which requires a UV laser) and a combination such as Fluo-3 and Fura Red, which can be used in instruments limited to excitation at 488 nm. Fluo-3 (with maximal fluorescence at 530 nm) increases its fluorescence with increasing Ca++ concentration. Fura Red (maximal fluorescence at 650 nm) decreases its fluorescence with increasing Ca++ concentration. The ratio of Fluo-3 to Fura Red fluorescence can be used to follow changes in Ca++concentration over time.

4Spectral Flow Cytometry Data Analysis

Spectral flow cytometry is used to sample more parameters per single cell or particle than traditional flow cytometry. In contrast to conventional flow cytometers, spectral flow analyzers and sorters substitute mirrors, optical filters, and photomultiplier tubes (PMTs) with a spectrograph and multichannel detector to use fluorescence or Raman spectroscopy.

FCS Express provides the capability to read raw spectral data from Cytek and Sony instruments in the 1D plot type, Spectrum Plots, for the analysis of spectral data. Data can be presented in the spectrum plot with numerous density, line, and backgating options to optimize displays for analysis. The color of each line can reflect the color of the gate in which the event falls effectively providing backgating information for spectral data. Multiple populations or files may also be overlaid with ease.