Spectrum 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 has the capability to read and display raw spectral data from Cytek and Sony instruments through Spectrum Plots.  Spectral data plotted in Spectrum plots links directly the standard flow cytometry single cell measurements to allow seamless transitions between standard flow cytometry analysis and spectral cytometry analysis.

 

Supported Data Formats:

 

Sony Instruments:

oSpectral data is stored with the .SRAW file format with both an .SRAW and .FCS file being produced. To analyze data from Sony instruments in spectrum plots the .sraw and .fcs files produced by the instrument must be placed in the same source folder. The names of the .SRAW and .FCS files must match.

 

 

oCytek Instruments: Spectral data is stored with the standard .FCS file format however two files will be produced, an "unmixed" data file and a "mixed" data file. The unmixed data file will have the same name as the mixed data file except it will use a suffix of "Unmixed" in the data file name. For instance:

 

 sample1.fcs and sample1_Unmixed.fcs

 

The file containing the suffix of "Unmixed" contains the FCS file data that was unmixed via the Cytek system while the file with the standard file name remains unmixed.
To view spectrum plots for the unmixed data set both data files most be placed in the same source folder with the names matching except for the "Unmixed" suffix.
To view spectrum plots for the mixed data set, only the original file without the "Unmixed" suffix must be present.