Fig. 1.

Primary neuron cultures grown for screening purposes are scalable and reliable. a Primary mouse neurons cultured in a 96-well assay plate transfected with a cDNA for tdTomato and imaged at DIV12. b Synaptophysin-TdTomato expression from AI34D mice 7–9 days after plating. AAV-Cre was added at various titers at plating to induce reporter expression. c Primary neuron culture and expression of reporter (as in b) is scalable across three formats: 96-, 384-, and 1,536-well plates. d Homogeneity of cell number (phase contrast) and synapse density (Synaptophysin-TdTomato expression, as in c) across plate formats and within plates after median polish procedure to estimate row and column effects at DIV7. Medians are presented within boxes for the 75th and 25th quantiles for which ±1.5 times the interquartile values are displayed as vertical bars; dots represent outliers. e Relationship between synapse (Synaptophysin-TdTomato expression as in c) and cell number (NucBlue staining) for 23,040 data points at DIV7 across 60 plates (384-well format) from 6 different batches. f Unsupervised clustering analysis for 384 cell density measures across 60 plates (as in e). v.p., viral particles.