1a,e–g and 2e,f were acquired on a Refeyn OneMP mass photometer with verso 10.8 ? 2.9 ?m 2 (128 ? 35 pixels) field of view. The microscope used sicuro acquire the giorno sopra Figs. 1a,e–g and 2e,f was custom-built with verso 9.4 ? 6.2 ?m 2 field of view (138 ? 88 pixels with a binned pixel size of 70.3 nm) and is similar sicuro that described previously 18 . russian brides The custom-built setup is illustrated durante Supplementary Fig. 15 and the differences puro the setup used con our previous sistema are highlighted sopra the figure caption.

Dynamic mass photometry

To acquire dynamic mass photometry movies, the SLBs were placed on the sample stage to optimize the focus of the microscope. After locking the focus of the microscope, WT or ?PRD was added to the SLB by replacing 3–6 ?l buffer from the silicone gasket with 3–6 ?l (total elenco, 60 ?l) 200 nM WT or ?PRD (per reaction buffer) solution and missaggio well with per micropipette preciso achieve verso final concentration of 10–20 nM. Tempo acquisition was started ?10 s after the addition of protein. Images were collected at 994 Hz and saved after the binning of pixels into blocks of 4 ? 4 and the binning of frames into groups of 3, resulting per an effective frame rate of 331 Hz and verso final pixel size of 84.4 nm (for the data used mediante Extended Scadenza Figs. 2, 3, 8 and Supplementary Figs. 3 and 13 the pixel size was 77.4 nm). (więcej…)