Unmix Using Weighted Least Squares — unmix.wls • AutoSpectral

This function performs unmixing of raw data using weighted least squares, AKA WLS, based on the provided spectra. Weighting is by channel power.

Usage

unmix.wls(raw.data, spectra, weights = NULL, noise.floor = 125)

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.
weights: Optional numeric vector of weights, one per fluorescent detector. Default is NULL, in which case weighting will be done by channel means.
noise.floor: Numeric, default 125. Lower clamp on mean channel signal before inversion when weights = NULL, preventing near-dark channels from producing an unbounded weight. Signal units, same convention as noise.floor elsewhere in the package.

A matrix containing unnmixed data with cells in rows and fluorophores in columns.