Fig. 2: Comparison of dye diffusibility into HM20 and K11M resins.
From: Visualizing suborganellar lipid distribution using correlative light and electron microscopy
a, The workflow for testing the penetration depth of the click reaction mixture into resins. Whole resin blocks were stained and subsequently sectioned vertically to assess penetration depth. b, Representative images of stained and vertical sectioned blocks of HM20 (upper) and K11M (lower), of samples loaded with (+lipid +UV) and without (−lipid −UV) PC(16:0|Y). Scale bars, 10 µm (overview images) and 1 µm (ROIs). The dotted red lines represent cell borders if not visible otherwise. c, The average line profile (indicated in b) of the fluorescence signal of vertical sections of HM20 and K11M with (black line) and without (dotted line) PC(16:0|Y). The error of the average line profile is shaded in grey and given as the confidence interval of 95%. The full width at half maximum (± s.d.) for the lipid-loaded samples is indicated. The purple bar indicates the mean resolution limit of the optical system as determined with 100-nm beads. Two independent samples were analysed for each resin, n (K11M sections) = 29 and n (HM20 sections) = 29. d, The workflow for testing the penetration depth of the click reaction mixture into resins. Whole resin blocks were stained and subsequently sectioned horizontally to assess the penetration depth. e, Representative images are shown of the first six horizontally cut sections merged and the first four sections separately of stained blocks for HM20 and K11M. Scale bar, 10 µm. The colours indicate the number of the section, with section 1 being the first section collected from the block. Section thickness, 100 nm. f, The Pearson coefficients between horizontal serial sections were determined. Coefficients were calculated pairwise for subsequent sections. Two independent samples were analysed for each resin and each analysis (n) comprises a ROI that contains at least one whole cell, n (K11M ROI) = 10 and n (HM20 ROI) = 9. The mean and s.e.m. are indicated as solid circles and error bars, respectively. Single data points are shown as transparent circles. High Pearson coefficients indicate an overlap of signal between sections, whereas low coefficients indicate mutually exclusive signals.
