qFlow · workbench
qFlow: a quantitative flow cytometry workbench
Flow cytometry is how Cleavr checks whether an RNA-level subtype pattern actually holds at the protein level, the thing that determines whether a receptor is usable for targeting. qFlow bundles the whole quantitative-flow toolchain into one page: turn raw fluorescence into an absolute receptor count with calibration beads, convert between standardized units, gate populations, score staining quality, compensate spillover, and read raw FCS files. It replaces round-tripping through a spreadsheet or several single-purpose calculators.
Absolute receptor counts require a fitted calibration curve. Rows without one are excluded and flagged, never approximated from a raw MFI/isotype ratio, which has no absolute meaning on its own. The receptor-quantification and subtype logic are identical to receptor_quantification.py and flow_cytometry_validation.py, so the browser and the scripts return the same numbers.
01Receptor quantification
Fit a per-date calibration curve (linear or log-log), convert MFI to absolute antibody binding capacity (receptors/cell), and compare the protein-level subtype pattern against the RNA-level screen.
02Unit conversion
Convert MFI to MESF, ERF (NIST-traceable), or PE molecules / ABC via QuantiBRITE-style beads, with F/P ratio handling and batch conversion.
03Gating & %positive
Compute percent-positive, median MFI, geometric mean, and robust SD from event-level data. Set a gate manually or derive it from a negative control.
04Stain / separation index
Score how cleanly a positive population separates from background, the standard metric for antibody titration and fluorochrome comparison.
05Compensation
Enter a spillover matrix and compensate measured values by inverting it: corrected = measured × spillover⁻¹.
06FCS import
Parse FCS 3.0 / 3.1 list-mode files in the browser (float, double, and integer data types), inspect parameters, and push any channel to the gating module.
6 modules
One page, local only
96 assertions
Automated test suite
Parity-checked
Vs. the Python scripts
10 references
Cited methodology
Lab tool · Browser-local