fig1

Figure 1. Light scatter-based analysis of buffer control and a mixed population of non-fluorescent SiNPs across platforms. Dot plots displaying the number of events as indicated in buffer control and the sample containing four size-defined populations of non-fluorescent SiNPs (68, 91, 113, 155 nm in diameter). Samples were measured on the NanoFCM with (A) an SSC (488 nm) threshold, on the IF with (B) an SSC (488 nm), on the CF with (C) an SSC (488 nm), on the IF with (D) an rw-FSC (488 nm) threshold, and on the CF with (E) a VSSC (405 nm) threshold. SiNP samples were pre-diluted shortly before measurements (1:100 for NF and 1:1,000 for both IF and CF). Samples were first gated to minimize the inclusion of doublets and aggregates (shown in Supplementary Figure 1). Next, each SiNP subset was gated, and the respective size is indicated within the plot; (F-I) Overlay histograms showing buffer control and gated SiNP subsets displaying each indicated scatter parameter. Identical buffer control and SiNP samples were acquired simultaneously on all three instruments in the same room under the same conditions. The figure shows data obtained from one selected sample dilution measured on each instrument with the indicated threshold (SSCt on NF, SSCt and FSCt on IF, and SSCt and VSSCt on CF). In total, 6 serial sample dilutions from the same stock solution were measured on the NF, while 12 serial dilutions were measured on the IF and the CF. CF: CytoFLEX LX; IF: BD Influx; NF: NanoFCM; FSC: forward scatter; rw-FSC: reduced wide-angle forward scatter; FSCt: forward scatter threshold; SSC: side scatter; SSCt: side scatter threshold; VSSC: violet side scatter; VSSCt: violet side scatter threshold; SiNPs: silica nanoparticles.