Highly multiplexed tissue imaging using repeated oligonucleotide exchange reaction
Abstract
Multiparameter tissue imaging enables analysis of cell‐cell interactions in situ, the cellular basis for tissue structure, and novel cell types that are spatially restricted, giving clues to biological mechanisms behind tissue homeostasis and disease. Here, we streamlined and simplified the multiplexed imaging method CO‐Detection by indEXing (CODEX) by validating 58 unique oligonucleotide barcodes that can be conjugated to antibodies. We showed that barcoded antibodies retained their specificity for staining cognate targets in human tissue. Antibodies were visualized one at a time by adding a fluorescently labeled oligonucleotide complementary to oligonucleotide barcode, imaging, stripping, and repeating this cycle. With this we developed a panel of 46 antibodies that was used to stain five human lymphoid tissues: three tonsils, a spleen, and a LN. To analyze the data produced, an image processing and analysis pipeline was developed that enabled single‐cell analysis on the data, including unsupervised clustering, that revealed 31 cell types across all tissues. We compared cell‐type compositions within and directly surrounding follicles from the different lymphoid organs and evaluated cell‐cell density correlations. This sequential oligonucleotide exchange technique enables a facile imaging of tissues that leverages pre‐existing imaging infrastructure to decrease the barriers to broad use of multiplexed imaging.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Mar 10, 2021
- Source ID
- 10.1002/eji.202048891
Entities
People
- Christian Schürch
- Garry P. Nolan
- Graham L. Barlow
- Gustavo Vazquez
- John Hickey
- Julia Kennedy‐darling
- Nikolay Samusik
- Salil S. Bhate
- Sarah Black
- Vishal G. Venkataraaman
- Yury Goltsev
Organizations
- Becton Dickinson
- Cancer Research UK
- Food and Drug Administration
- Gates Foundation
- National Institutes of Health
- Silicon Valley Community Foundation
- Stanford University
- Swiss National Science Foundation
- United States Department of Defense