Journal: STAR Protocols
Article Title: Protocol for iterative indirect immunofluorescence imaging of frozen mouse intestinal tissues
doi: 10.1016/j.xpro.2026.104410
Figure Lengend Snippet: Multiplexed imaging of protein targets from the same tissue sample using Cryo-4i Representative images showing the expression patterns of different protein targets across five rounds of 4i: (A) phospho-Rb (round 1), (B) phospho-Smad1/5/8 (round 2), (C) Vimentin (round 3), (D) Mucin 2 (round 4), (E) Aldolase B (round 5), and (F) merged image of all rounds. All targets were detected using primary antibodies raised in rabbits. For nuclear segmentation, H2B-mScarlet (from a fluorescent reporter mouse) was used in round 1, while Histone H3 and E-cadherin were used from round 2 onwards. Representative data from three biological replicates (n = 3 mice). Scale bars, 50 μm.
Article Snippet: Phospho-Smad1/5/8 (Ser463/465) (dilution 1:50) , Merck , Cat#AB3848-I; RRID: AB_177439.
Techniques: Imaging, Expressing