AutoSpill

AutoSpill computes spillover coefficients and spectral signatures for fluorophores in fluorescence-based high-parameter flow cytometry to enable accurate compensation, unmixing, and autofluorescence correction.

Key Features:

• Automated Gating: Performs automated gating of cells.
• Initial Spillover Matrix Calculation: Computes an initial spillover matrix using robust linear regression.
• Iterative Refinement: Iteratively refines spillover coefficients to minimize compensation errors.
• Autofluorescence Compensation: Compensates for autofluorescence by treating it as an endogenous dye in unstained controls.
• Simplified Control Preparation: Operates without requiring well-defined positive and negative populations for single-color controls.
• Automatic Tuning of Spillover Matrix: Automatically computes tuning of the spillover matrix.
• AutoSpread Integration: Integrates with AutoSpread to provide a robust estimate of the Spillover Spreading Matrix (SSM).

Scientific Applications:

• Compensation and Unmixing: Compensation and unmixing of high-parameter flow cytometry fluorescence data.
• Autofluorescence Correction: Correction of autofluorescence in fluorescence-based systems using unstained controls.
• SSM Estimation: Estimation of the Spillover Spreading Matrix (SSM) via AutoSpread for high-parameter cytometry.
• Robust Compensation Workflows: Support for robust compensation workflows in modern high-parameter flow cytometry experiments.

Methodology:

Computational steps include automated gating, robust linear regression to compute an initial spillover matrix, iterative refinement of spillover coefficients, treating autofluorescence as an endogenous dye using unstained controls, automatic tuning of the spillover matrix, and use of AutoSpread to estimate the Spillover Spreading Matrix (SSM).