Figure 2
From: Simultaneous expansion microscopy imaging of proteins and mRNAs via dual-ExM
Effect of a post-fixation step between IF and FISH in the IF-FISH process. (a,b) Maximum intensity projection (MIP) of a z-stack image of NIH-3T3 cells labeled with an antibody against vimentin (red) and DAPI (blue). After antibody labeling, cells were either post-fixed or not post-fixed and then proceeded to FISH. (a) Cells post-fixed after IF. Vimentin signals are visible. (b) Cells not post-fixed after IF. Most of the vimentin signals were lost. (c) Relative fluorescence intensity of vimentin fibers of the cells shown in (a) and (b). (d,e) Validation of the mRNA labeling of post-fixed cells. (d) MIP image of a NIH-3T3 cell, labeled with an RNAscope probe against Gapdh mRNA (green) and antibody against vimentin (red). (e) MIP image of NIH-3T3 cells, labeled with an RNAscope negative control probe (DapB) and antibody against vimentin. Scale bars in (a,b) 50 μm; (d,e) 10 μm.
