Fig 1. Study design.

A) Each kit includes all consumables and reagents required for the biocytometry protocol. These comprise the bioparticle reagent, a set of HUMO samples (negative, low and high), a hydrogel-based incubation medium, resuspension and luminescence buffers, a luminescence substrate, evaluation strips and a plate adapter for the final luminescence readout in a microplate reader. B) HUMO samples of 100,000 cells were provided with each biocytometry kit. The concentrations of targets in the three HUMO samples were: 0 in 100,000 cells (negative), 1 in 100,000 cells (low), and 35 in 100,000 cells (high). For measurement, 56,000 cells were analyzed across 8 wells. The figure illustrates how signal-to-noise ratio values and the target count estimates are proportional to, and derived from, the luminescence readout of individual sample wells. C) Distribution of all 84 assays from the study dataset, 78 assays encompassing HUMO samples of every type (negative, low and high) complemented by 6 complementary assays from the inter-user reproducibility study encompassing high HUMO samples only. The figure outlines the distribution of the biocytometry kits and HUMO samples and the number of assays carried out by various laboratories.