Highly Sensitive and Multiplexed Protein Imaging With Cleavable Fluorescent Tyramide Reveals Human Neuronal Heterogeneity
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 si...
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doaj-2af2b19b3b9f4b0196c274692af841a62021-01-08T04:52:29ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2021-01-01810.3389/fcell.2020.614624614624Highly Sensitive and Multiplexed Protein Imaging With Cleavable Fluorescent Tyramide Reveals Human Neuronal HeterogeneityRenjie Liao0Manas Mondal1Christopher D. Nazaroff2Christopher D. Nazaroff3Diego Mastroeni4Diego Mastroeni5Paul D. Coleman6Paul D. Coleman7Joshua Labaer8Jia Guo9Biodesign Institute & School of Molecular Sciences, Arizona State University, Tempe, AZ, United StatesBiodesign Institute & School of Molecular Sciences, Arizona State University, Tempe, AZ, United StatesBiodesign Institute & School of Molecular Sciences, Arizona State University, Tempe, AZ, United StatesDivision of Pulmonary Medicine, Department of Biochemistry and Molecular Biology, Mayo Clinic Arizona, Scottsdale, AZ, United StatesArizona State University-Banner Neurodegenerative Disease Research Center, Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ, United StatesL.J. Roberts Center for Alzheimer's Research, Banner Sun Health Research Institute, Sun City, AZ, United StatesArizona State University-Banner Neurodegenerative Disease Research Center, Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ, United StatesL.J. Roberts Center for Alzheimer's Research, Banner Sun Health Research Institute, Sun City, AZ, United StatesBiodesign Institute & School of Molecular Sciences, Arizona State University, Tempe, AZ, United StatesBiodesign Institute & School of Molecular Sciences, Arizona State University, Tempe, AZ, United StatesThe 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 co-regulated proteins. We also profiled >6,000 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 sub-regions of the hippocampus consist of neurons from distinct clusters.https://www.frontiersin.org/articles/10.3389/fcell.2020.614624/fullimmunohistochemistryimmunofluorescencesingle-cellspatial proteomicshippocampus |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Renjie Liao Manas Mondal Christopher D. Nazaroff Christopher D. Nazaroff Diego Mastroeni Diego Mastroeni Paul D. Coleman Paul D. Coleman Joshua Labaer Jia Guo |
spellingShingle |
Renjie Liao Manas Mondal Christopher D. Nazaroff Christopher D. Nazaroff Diego Mastroeni Diego Mastroeni Paul D. Coleman Paul D. Coleman Joshua Labaer Jia Guo Highly Sensitive and Multiplexed Protein Imaging With Cleavable Fluorescent Tyramide Reveals Human Neuronal Heterogeneity Frontiers in Cell and Developmental Biology immunohistochemistry immunofluorescence single-cell spatial proteomics hippocampus |
author_facet |
Renjie Liao Manas Mondal Christopher D. Nazaroff Christopher D. Nazaroff Diego Mastroeni Diego Mastroeni Paul D. Coleman Paul D. Coleman Joshua Labaer Jia Guo |
author_sort |
Renjie Liao |
title |
Highly Sensitive and Multiplexed Protein Imaging With Cleavable Fluorescent Tyramide Reveals Human Neuronal Heterogeneity |
title_short |
Highly Sensitive and Multiplexed Protein Imaging With Cleavable Fluorescent Tyramide Reveals Human Neuronal Heterogeneity |
title_full |
Highly Sensitive and Multiplexed Protein Imaging With Cleavable Fluorescent Tyramide Reveals Human Neuronal Heterogeneity |
title_fullStr |
Highly Sensitive and Multiplexed Protein Imaging With Cleavable Fluorescent Tyramide Reveals Human Neuronal Heterogeneity |
title_full_unstemmed |
Highly Sensitive and Multiplexed Protein Imaging With Cleavable Fluorescent Tyramide Reveals Human Neuronal Heterogeneity |
title_sort |
highly sensitive and multiplexed protein imaging with cleavable fluorescent tyramide reveals human neuronal heterogeneity |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Cell and Developmental Biology |
issn |
2296-634X |
publishDate |
2021-01-01 |
description |
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 co-regulated proteins. We also profiled >6,000 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 sub-regions of the hippocampus consist of neurons from distinct clusters. |
topic |
immunohistochemistry immunofluorescence single-cell spatial proteomics hippocampus |
url |
https://www.frontiersin.org/articles/10.3389/fcell.2020.614624/full |
work_keys_str_mv |
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