Figure 5

PALM measurement precision and quantification of Na,K-ATPase in spines. (a) Histogram of unfiltered detection precision for single recordings of PAGFP-ATP1a1 or mEos3.2-ATP1a1 in hippocampal cells. Typical mean precision was estimated at 25 nm and 20 nm for PAGFP and mEos3.2, respectively. (b) Example of single spine PALM image with detected localization for mEos3.2-ATP1a3. Scale bar = 200 nm. (c) Curve fitting according to Annibale et al. to estimate the overcounting due to photoblinking in the fast time regime (fitting done with first 10 frames). (d) Boxplots of total molecular density in spines comparing PAGFP and mEosFPs for each of the Na,K-ATPase isoforms. Boxes present the interquartile range of each group and median value (red line). Significance test shows no difference of median values between different fluorescent proteins (Mann–Whitney U-test).