Fig. 2: Ultrastructural staining on BOOST processed samples.
From: BOOST: a robust ten-fold expansion method on hour-scale
A Schematic of ultrastructural staining after BOOST denaturation. B Comparison of the microwave-assisted NHS ester staining time with ExM techniques using post-processing staining. C 3D visualization of a mitotic U2-OS cell, EF = 10.2 ×. D Left, stained U2-OS cell, EF = 10.2 ×. Blue arrow, granular component of nucleolus. Line profile along the dashed line indicating a partially complementary staining pattern of NHS ester and SYTOX Green in the nucleolus. E BOOST preserves ultrastructural details in vibratome-cut mouse tissue sections. (i) Synaptic structures in mouse brain section, EF = 9.5 ×. (ii) Mouse sperm annulus (green arrowhead), midpiece (below the annulus), and principal piece (above the annulus) in the testicle, EF = 9.4 ×. (iii) Mouse hepatocytes showing endoplasmic reticulum (purple arrowhead), mitochondria (blue arrowhead), and nuclear pores (green arrowhead) in the liver, EF = 9.1 ×. (iv) Mouse cardiomyocyte showing densely labeled A-band with intercalated M-line (green arrowheads) in heart, EF = 8.9 ×. (v) Mouse podocyte foot process (green arrow) and continuous globular basement membrane (blue arrow) in kidney with zoomed-in view (right), EF = 9.0 ×. F NHS ester stained mouse tumor-bearing lung FFPE section before and after BOOST treatment. Post-BOOST images were taken at ~ 2.6 × shrunken state in PBS. Magnified views showing the tumor before and after expansion. G NHS ester stained human prostate cancer FFPE section before and after BOOST processing. Magnified views showing resolved tumor cells after expansion, EF = 8.7 ×. H Visualization of mitochondria (green arrowheads) in the FFPE human prostate cancer section, EF = 8.7 ×. Additional note: graphical icons in 2A are created using Biorender. Scale bars (biological scale): 2 μm (C); 2 μm (D, left); 1 μm (D, right); 2 μm (E, left and top right); 0.2 μm (E(i), all four panels); 0.2 μm (E(ii)); 1 μm (E(iii), top and bottom); 0.5 μm (E(iv)); 2 μm (E(v), left); 0.5 μm (E(v), right); 1 mm (F, top left and top middle); 400 μm (F, bottom left and bottom middle); 500 μm (G, left and right normal views), 20 μm (G, left and right boxed zoomed-in views); 1 μm (H).
