Abstract
Traditional immunofluorescence (IF) imaging assays are limited to the detection of just a few markers due to spectral overlap of fluorescent emission bands. Furthermore, standard fluorescent imaging instruments have a dynamic range that is too narrow to capture the full range of expression values seen in biology, precluding the accurate quantification of single-cell target expression. Here we describe a protocol for detection and quantification of dozens of protein targets with single-cell quantitative precision using an iterative staining approach called ChipCytometry™.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Editorial (2022) Catching up with multiplexed tissue imaging. Nat Methods 19:259. https://doi.org/10.1038/s41592-022-01428-z
Jarosch S, Kohlen J, Wagner S et al (2021) Multiplexed imaging and automated signal quantification in formalin-fixed paraffin-embedded tissues by ChipCytometry. Cell Rep Methods 1:100104. https://doi.org/10.1016/j.crmeth.2021.100104
FitzPatrick MEB, Provine NM, Garner LC et al (2021) Human intestinal tissue-resident memory T cells comprise transcriptionally and functionally distinct subsets. Cell Rep 34:108661. https://doi.org/10.1016/j.celrep.2020.108661
Lett MJ, Mehta H, Keogh A et al (2022) Stimulatory MAIT cell antigens reach the circulation and are efficiently metabolised and presented by human liver cells. Gut:1–13. https://doi.org/10.1136/gutjnl-2021-324478
Schupp J, Christians A, Zimmer N et al (2021) In-depth immune-oncology studies of the tumor microenvironment in a humanized melanoma mouse model. Int J Mol Sci 22(3):1011. https://doi.org/10.3390/ijms22031011
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Campbell, T.D., Christians, A., Tyler, M.A. (2023). Highly Multiplexed Immunofluorescence Imaging for Quantitative Spatial Analysis in Tissue Samples with ChipCytometry™. In: Kalyuzhny, A.E. (eds) Signal Transduction Immunohistochemistry. Methods in Molecular Biology, vol 2593. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2811-9_21
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2811-9_21
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2810-2
Online ISBN: 978-1-0716-2811-9
eBook Packages: Springer Protocols