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Table 1 Summary of marker optimizations and results

From: Fluorescence/luminescence-based markers for the assessment of Schistosoma mansoni schistosomula drug assays

Marker Type Marker Assay Optimal incubation time S:B ratio ≥ 3:1? Correlation to NTS viability/cytotoxicity Selected for drug assay testing? Justification
Viability Markers CellTiter-Glo® 15 min Yes, with ≥ 100 NTS Strong Yes Met criteria
  Resazurin 24 hours Yes, with ≥ 200 NTS Strong Yes Met criteria
Cytotoxicity Markers Vybrant® 70 min Almost, with ≥ 300 NTS Strong up to 300 NTS Yes Met criteria
  CytoTox-ONE 2 hours No Strong No S:B ratio low, large standard deviations between data points
  CellTox™ Green Cytotoxicity Assay 24 hours Yes, with ≥ 400 NTS Strong with ≥ 400 NTS No Viable NTS died within 24 hours of exposure to dye; strong signals from completely lysed cells only
Multiplex Markers LIVE/DEAD® Viability/Cytotoxicity Kit Does not exist No Poor for both live and dead NTS No Poor correlation to NTS viability/cytotoxicity
  ApoTox-Glo™ Does not exist No Strong for live-cell marker, poor for dead-cell marker No Markers induced spazzing and death of NTS after 6 hours
Experimental Markers OmniCathepsin™ 2 hours (with 10 μM marker concentration) No Good: Differential signals for live vs. dead observed No S:B ratio too low, not enough difference between live and dead NTS
  FluoForte® Calcium Assay 1.5 hours with ≥ 200 dead NTS Yes, with ≥ 200 dead NTS (lysed only) Poor: strong staining for completely lysed cells only No Stains only completely lysed cells
  DAPI 15 min (with 10 μg/ml dye concentration) No Good: stained many dead NTS phenotypes No Background fluorescence too high, signals too low
  Hoechst 33258 15 min (with 1 μg/ml dye concentration) No Good: stained many dead NTS phenotypes No Background fluorescence too high, signals too low