Extended Data Fig. 6: A comparison of the performance of MTn tag labeling, Tokuyasu technique, and conventional immuno-EM method.
From: Genetically encoded tags for direct synthesis of EM-visible gold nanoparticles in cells
a, b, Expression patterns of the ER-located membrane proteins, Ost4-GFP and Ost4-GFP-MTn, in S. pombe observed by live-cell fluorescence imaging. c, Immunogold staining of Ost4-GFP in yeast cell using anti-GFP (goat) antibody (Cat. No.600-101-215, Lot.35059, Rockland) and protein A-10nm gold particles (Cat.No.25285, Code:110.111, EMS) on a cryosection prepared with Tokuyasu technique. d, Direct synthesis of AuNPs on Ost4-GFP-MTn in a yeast cell using Scheme 2b (PIPES) for ANSM AuNP synthesis. e, Immunogold staining of Ost4-GFP using anti-GFP (Rabbit) antibody (Cat. No. ab6556, Lot.GR3252667-1, Abcam) and anti-rabbit IgG antibody-10nm gold particles (Cat.No.G7402-.4 ml, Lot.SLBT0649, Sigma) on LR white embedded plastic section of a yeast cell (cell prepared with HPF, FSF with 0.1% UA + 5% H2O in acetone). f, Immunogold staining of Ost4-GFP-MTn in yeast cell (the same experimental setting as used for e, except embedded in HM20 resin). Notably, the labeling density with ANSM AuNPs synthesis on Ost4-GFP-MTn (d) is ~10-20x higher than those with immunogold staining (c, e, f). Representative images were selected from n = 3 (a, b, c, e, f), 5 (d) independent experiments with similar results.
