Fig. 2: Validation of Magnify in several tissue types.

a,b, Example of pre-expansion images of human kidney imaged at ×60 and processed with SOFI (a) compared with the same FOV postexpansion with Magnify taken at ×40 (b). Postexpansion images are maximum intensity projected over 25 frames in z. c–e, RMS length measurement error as a function of measurement length for pre-expansion versus postexpansion images for DAPI (c), ACTN4 (d) and Vimentin (e). Solid line, mean of channel; shaded area, s.e.m.; n = 5 technical replicates; average EF = 8.64× (s.e.m. 0.24). f,g, Example images of human prostate imaged as in a and b. Postexpansion images maximum intensity projected over three frames. h,i, RMS length measurement error as a function of measurement length for pre-expansion versus postexpansion images of DAPI (h) and ATPIF (i). Solid line, mean of channel; shaded area, s.e.m.; n = 4 technical replicates; average EF = 10.38× (s.e.m. 0.57). j–o, Validation of Magnify across several human tissue types. FFPE samples of human tissue were imaged at ×40 (top left). Images were taken at ×60 and processed with SOFI (bottom left). The white box indicates the FOV of the higher magnification images. The samples were then processed with the Magnify protocol, and the same FOVs were imaged postexpansion in water at ×10 (top right) and ×40 (bottom right). Postexpansion images were projected over 4–17 z slices. EFs in water were colon 8.85× (j), breast 9× (k), uterus 8× (l), placenta 8.75× (m), thymus 10.00× (n) and thyroid 10.59× (o). p–r, Example 3D images of human tissues: kidney (EF = 8.68×) (p), colon (EF = 9.67×) (q) and uterus (EF = 8×) (r). Dashed white boxes, zoomed in regions. Scale bars (yellow, postexpansion images). a, 5 μm; b, 5 μm (physical scale postexpansion, 40.75 μm; EF = 8.15×); f, 5 μm; g, 5 μm (physical scale postexpansion: 51.9 μm; EF = 10.38×); j–o, top, 10 μm; bottom, 1 μm; p–r, 5 μm. Scale bars are all in biological scale.