Extended Data Fig. 1: Parameter optimizations of sample preparation.
From: Electrospray-assisted cryo-EM sample preparation to mitigate interfacial effects
a, Structural preservation of apo-ferritin under different spraying conditions. Several parameters, like sample flow rate, high voltage, and protein concentration, affect protein samples of intact states differently. The particles in the blue squares represent the shape of the intact protein, while the particles in the green squares represent the disrupted and dispersed state. Conditions with moderately medium flow rates, lower voltages, and higher sample concentrations contribute to better structural integrity. Metal materials with low conductivity in nano-ESI would also make a difference in structural maintenance. b, shows the statistical analysis of the percentage of structurally intact proteins in EM negative-staining under different spraying conditions. Micrographs were collected and analyzed independently for each flow rate, high voltage value, protein concentration, and conductive material of nano-ESI. The sample sizes (n) were 5, 6, 15, and 5 for flow rates; 8, 17, and 7 for high voltage values; 15, 17, and 20 for protein concentrations; and 6, 12, 14, and 16 for conductive materials of nano-ESI, respectively. The bars indicating the highest proportions of intact particles represent the recommended optimal parameters. The central measure of the error bar is the mean, while the error bar represents the 95% confidence interval. The source data is included in a dedicated Source Data file.
