Highly accurate image registration for 3D multiplexed cyclic imaging using dense labeling in expandable tissue gels
Fig 5
Registration of multiplexed images via actin staining in expanded cultured cells.
(a) Composite image generated by registering two images acquired from the first and second imaging rounds. Phalloidin-labeled actin fibers were used as fiducial markers. The yellow box displays six-color images, including the DAPI channel. Gray, nucleus; red, actin; cyan, vimentin; blue, laminA/C; yellow, CCP; and magenta, cytokeratin 8/18. (b–c) Magnified views of the boxed region in a. Scale bars: (a) 2 µm; (b–c) 500 nm. All length scales are presented in pre-expansion dimensions. Number of sample N = 1 acquired from a single substrate.