High-dimensional imaging of FFPE mouse liver tissues by mIF. (IMAGE)
Caption
(A) Sequential mIF simplified workflow to analyse FFPE liver samples. Panel A was created with BioRender.com. (B) An FFPE liver section from an adult healthy mouse was analysed using the mIF method. Typical output images after alignment are displayed. Areas of interest are delineated and shown in the following panels at a higher magnification. (C) Area 3 highlights a selected region of the liver and displays single channel as well as merged images and binary segmentation (lower image) of the two indicated markers. (D) Area 4 was used to demonstrate the possibility of using different sets of markers to highlight specific cellular contexts according to the researcher’s objectives. (E) Nuclear staining (DAPI) over six consecutive days of imaging and antibody stripping is shown individually and as a merged image for Area 5, as a quality control for tissue integrity. The following fluorophores were conjugated to the appropriate secondary antibodies: α-SMA: A488; CD3: A647; CK7: A750; CLEC4F: A647; HNF4α: A750; IBA1: A750; LYVE-1: A647; MPO: A555. Ab, antibody; DAPI, 4′,6-diamidino-2-phenylindole; FFPE, formalin-fixed paraffin-embedded; mIF, multiplex immunofluorescence.
Credit
By Bianca Franco Leonardi, Guo Yin, Natalja Amiridze, et al.
Usage Restrictions
Credit must be given to the creator.
License
CC BY