Supplementary Figure 9: Other sample orientations and alternative reduction-mediated fluorophore removal timeline for Expansion-SABER.

(a) Additional post-expansion images of neuronal synapses showing different orientations, as in Fig. 6b. (b) High-zoom comparison of Vimentin and Calretinin signals with and without expansion. The image with expansion is the overlay of the Vimentin and Calretinin images from Fig. 6c, whereas the image without expansion was derived from images in Fig. 5a. Scale bar indicates the physical size after expansion (~3-fold). (c) Alternative fluorophore removal strategy using TCEP reduction in thick expanded samples using TCEP. A mouse retina section stained with SV2 was expanded and visualized with disulfide bond modified imager strands. The fluorophores (Alexa 647) on the imager strands were cleaved using TCEP reduction. The fluorescence signal was monitored using a confocal microscope in a time course for 10 min. (d) Quantification of fluorescence signals in c before and after TCEP reduction.