'Iterative Bleaching Extends Multiplexity' facilitates simultaneous identification of all major retinal cell types.

ABSTRACT: To understand the multicellular composition of tissues, and how it is altered during development, ageing and/or disease, we must visualise the complete cellular landscape. Currently, this is hindered by our limited ability to combine multiple cellular markers. To overcome this, we adapted a highly multiplexed immunofluorescence (IF) technique called 'Iterative Bleaching Extends Multiplexity' (IBEX) to the zebrafish retina. We optimised fluorescent antibody micro-conjugation to perform sequential rounds of labelling on a single tissue to simultaneously visualise all major retinal cell types with 11 cell-specific antibodies. We further adapted IBEX to be compatible with fluorescent transgenic reporter lines, in situ hybridisation chain reaction (HCR), and whole-mount immunofluorescence (WMIF

SUBMITTER: Kothurkar AA 

PROVIDER: S-EPMC11827602 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature