Assign AF Spectrum By Fluorophore Projection
Source:R/assign_af_fluorophores.R
assign.af.fluorophores.RdProjects the autofluorescence spectral variants into fluorophore unmixed space to determine which best fits each cell (event). A fast approximation for brute force sequential unmixing method in early versions of AutoSpectral. Provides essentially identical results to minimization of fluorophore signal (dist0 method). Substantially faster. Use L1 (absolute value) minimization, which works better than the standard L2 (squared error).
Arguments
- raw.data
Expression data from raw FCS files. Cells in rows and detectors in columns. Columns should be fluorescent data only and must match the columns in spectra.
- spectra
Spectral signatures of fluorophores, normalized between 0 and 1, with fluorophores in rows and detectors in columns.
- af.spectra
Spectral signatures of autofluorescences, normalized between 0 and 1, with fluorophores in rows and detectors in columns.