Optimization of fixed-permeabilized cell monolayers for high throughput micro-neutralizing antibody assays: application to the zebrafish/viral hemorrhagic septicemia virus (vhsv) model.

•A new micro-neutralization high-throughput centrifugation-free method is described.•Host cell monolayers were fixed to poly-d-Lys coated solid-phases.•Saponin-containing buffers were used for immunostaining.•Cells were detached by trypsin digestion for batch flow cytometry.•Viral-neutralizing antibodies were successfully assayed in plasma from VHSV-immunized zebrafish (Danio rerio).