Supplementary Figure 3: Imaging fluorescent nanobeads.
From: An acquisition and analysis pipeline for scanning angle interference microscopy
(a) SAIM prediction for 200 nm fluorescent nano-sized beads. Model of the SAIM intensity function a 200nm sphere, stained throughout generated by dividing the sphere in 9 parts, calculating the SAIM curve for each part, and averaging the predicted intensity for the 9 parts at each angle (weighted by the fraction of the sphere's volume contained in each part) (blue line), compared to the SAIM curve of a point source at the radius of the bead (orange line). (b) Histogram of fluorescent nanobead radii measured by SAIM. Left, histogram of yellow-green nanobeads. Measured heights of nominal “20 nm” radius (company provided specifications) nanobeads shown in light green (n = 1330, mean = 33 ± 13 nm), and nominal “50 nm” radius nanobeads shown in dark green (n = 823, mean = 63 ± 11 nm). Right, histogram of red nanobeads. Measured heights of nominal 20 nm radius nanobeads shown in light red (n = 451, mean = 28 ± 8 nm), and nominal 50 nm radius nanobeads shown in dark red (n = 1391, mean = 64 ± 7 nm). (c) Comparison of nanobead radius measured by SAIM or electron microscopy. Color-coding corresponds to panel b, radii measured by SAIM are solid colored, radii measured by electron microscopy are indicated by hatch marks. Error bars denote standard deviation.
