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The testing was performed according to Zucker, 2001 Fluorescent plastic slides were supplied by Chroma.
Apo TIRF 60x N.A. 1.49 objective was tested with the three laser lines (405,
488, 543 nm). Full field of view (212x212 µm resolved 512x512 px) was
evaluated. 35 Z-stacks (10 µm thick, step size 1 µm, middle point
reference 25 µm in the slide, 0 set to correspond to maximum primary reflectance
by the slide) were acquired at each wavelength. Corresponding Z slices (10 overall
for each laser line) were averaged and the diagonal intensity profiles plotted
as depicted on the images.
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