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. 2020 Sep 15;11:4632. doi: 10.1038/s41467-020-18422-8

Fig. 1. miriEx robustly preserves antigens across multiple rounds of immunostaining, imaging, and stripping.

Fig. 1

a miriEx strategy to simultaneously measure molecular profile, morphology, and/or connectivity. Tissue samples are embedded in a hydrogel to create expandable gel-tissue hybrids. The sample can then undergo multiple rounds of immunostaining, imaging, and stripping to measure various neuron properties. We then register and merge the different rounds of imaging to correlate the results. b Five distinct rabbit antibodies were used across 7 sequential rounds of immunostaining, with PV re-probed in rounds 4 and 7 to demonstrate retention of antigen. The merged image shows r1 PV, r2 Calb, r3 Cb1R, r5 NOS, and r6 SERT. c Signal to noise was quantified for PV immunostaining in round 1, round 4, and round 7 (n = 10 neurons). Abs antibodies, PV parvalbumin, Calb calbindin, Cb1R cannabinoid receptor type 1, Nos nitric oxide synthase, SERT serotonin transporter. Scale bar: (b) 25 μm (pre-expansion size). Expansion factor: (b) ~2×. See Supplementary Table 1 for more details.