Multiplex Staining by Sequential Immunostaining and Antibody Removal on Routine Tissue Sections.

ABSTRACT: Multiplexing, labeling for multiple immunostains in the very same cell or tissue section in situ, has raised considerable interest. The methods proposed include the use of labeled primary antibodies, spectral separation of fluorochromes, bleaching of the fluorophores or chromogens, blocking of previous antibody layers, all in various combinations. The major obstacles to the diffusion of this technique are high costs in custom antibodies and instruments, low throughput, and scarcity of specialized skills or facilities. We have validated a method based on common primary and secondary antibodies and diffusely available fluorescent image scanners. It entails rounds of four-color indirect immunofluorescence, image acquisition, and removal (stripping) of the antibodies, before another stain is a

SUBMITTER: Bolognesi MM 

PROVIDER: S-EPMC5533273 | biostudies-literature | 2017 Aug

REPOSITORIES: biostudies-literature