RT Journal Article
SR Electronic
T1 Highly sensitive in situ proteomics with cleavable fluorescent tyramide reveals human neuronal heterogeneity
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 539106
DO 10.1101/539106
A1 Renjie Liao
A1 Manas Mondal
A1 Christopher D. Nazaroff
A1 Diego Mastroeni
A1 Paul D. Coleman
A1 Jia Guo
YR 2019
UL http://biorxiv.org/content/early/2019/02/08/539106.abstract
AB The ability to comprehensively profile proteins in intact tissues in situ is crucial for our understanding of health and disease. However, the existing methods suffer from low sensitivity and limited sample throughput. To address these issues, here we present a highly sensitive and multiplexed in situ protein analysis approach using cleavable fluorescent tyramide and off-the-shelf antibodies. Compared with the current methods, this approach enhances the detection sensitivity and reduces the imaging time by 1-2 orders of magnitude, and can potentially detect hundreds of proteins in intact tissues at the optical resolution. Applying this approach, we studied protein expression heterogeneity in a population of genetically identical cells, and performed protein expression correlation analysis to identify coregulated proteins. We also profiled >6000 neurons in a human formalin-fixed paraffin-embedded (FFPE) hippocampus tissue. By partitioning these neurons into varied cell clusters based on their multiplexed protein expression profiles, we observed different subregions of the hippocampus consist of neurons from distinct clusters.