Pangnostics dFC — See the 90% your cytometer is missing

The Pangnostics dFC is an ultra-high-sensitivity single-molecule counter in a familiar flow-cytometry format — counting, sizing, and phenotyping every extracellular vesicle, LNP, viral vector, and antibody that flows through, without a single calibration bead.

Key numbers: approximately 100% single-molecule detection efficiency; 4 lasers by 12 detection channels; 1 to 96 samples per run; 0 standard curves required.

Why it matters

For decades, flow cytometry has driven our understanding of cellular heterogeneity. But when the same instruments are turned on the nanoscale — extracellular vesicles, lipid nanoparticles, viral vectors — a large fraction of every sample stays hidden below the instrument's detection floor. Most EVs are smaller than 70 nm and dimmer than a handful of fluorophores, so they simply aren't counted. Every downstream conclusion inherits that blind spot.

Pangnostics dFC closes the gap with a fundamentally different optical design: planar microfluidics and line-confocal detection that count individual fluorophore-tagged molecules directly, with essentially 100% single-molecule detection efficiency. The result is absolute, calibration-free quantitation — and a per-particle view of size, surface markers, and copy number that bulk methods average away.

How digital flow cytometry works

• Flow — A 10 µL sample is drawn into a planar microfluidic chip and flows single-file through a detection channel just a couple of micrometres across, so particles pass the laser one at a time.
• Illuminate — Four spatially separated lasers (405, 488, 561 and 637 nm) are shaped into a line-confocal light sheet that spans the channel, defining a tiny, evenly-lit detection volume every particle must cross.
• Count — Each fluorophore that crosses the sheet emits a burst of photons recorded by single-photon detectors. Bursts above a noise threshold are counted as individual molecules, with essentially 100% detection efficiency and a very low false-positive rate.
• Phenotype — Twelve channels read the bursts across up to eighteen dyes at once. Colocalizing signals on the same particle gives a true multi-parameter phenotype — and the copy number of each marker, per vesicle.

Key capabilities — six things bulk methods can't do

• Single-molecule sensitivity — validated detection efficiency near 100% across more than 30 fluorescent dyes. Every photon-emitting molecule is counted, including the dimmest, not estimated from a curve.
• High-plex phenotyping — four lasers and twelve simultaneous channels resolve surface-marker combinations on individual nanoparticles in a single run, expandable for higher-plex assays.
• Absolute quantitation — direct digital counting yields true concentrations and biomarker copy numbers, with no calibration beads, no reference ladders, and no ~20% bead-calibration uncertainty.
• Flexible throughput — run a single urgent sample or a full 96-well plate on the same platform, with a familiar flow-cytometry workflow and no specialist operator required.
• Reveals heterogeneity — surfaces the dim, small, and rare subpopulations, well under 1% of total particles, that bulk and commercial high-sensitivity platforms miss by design.
• Reagent authentication built in — measures the absolute concentration of antibody-dye conjugates that can't be read at 280 nm, plus dye-to-protein ratio, removing lot-to-lot reagent variability as a source of irreproducibility.

Applications — from discovery to lot release

• Extracellular vesicles & exosomes — phenotype individual EVs across twelve colors; quantify tetraspanin and engineered-marker copy number and absolute subpopulation concentration for liquid biopsy and EV therapeutics.
• Lipid nanoparticles (LNPs) — characterize mRNA and gene-therapy LNPs at the single-particle level: payload distribution, surface composition, and encapsulation uniformity.
• AAV & viral vectors — distinguish full, empty, and partial capsids; measure titer and payload heterogeneity for vector development and release characterization.
• Protein aggregates & complexes — resolve oligomeric states and RNA-binding protein complexes that bulk methods cannot separate from vesicular populations.
• Analytical & therapeutic antibodies — authenticate dye-to-protein ratios, quantify aggregation, and verify conjugate concentration.
• Nanoparticle standardization — reference-grade absolute concentrations make dFC an orthogonal method for cross-laboratory harmonization and reagent QC.

Rigor & reproducibility

Absolute counting removes the calibration drift, curve-fitting, and vendor-dependent reagent variability that compromise quantitative EV flow cytometry. Copy-number and sizing methods have been validated against orthogonal techniques including super-resolution and single-molecule TIRF microscopy, and the platform is designed to support standardized community reporting. Aligned with MISEV2023 reporting and the MIFlowCyt-EV framework; orthogonally validated against super-resolution / TIRF; approximately 4 orders of magnitude dynamic range.

Contact Bioparticle

• Email: info@bioparticle.com
• Phone: +1-480-604-5662
• Address: 2820 N Torino Ave, Tucson, Arizona 85712, US

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