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McKinley ET, Sui Y, Al-Kofahi Y, et al. Optimized multiplex immunofluorescence single-cell analysis reveals tuft cell heterogeneity. JCI Insight. 2017;2(11)doi: 10.1172/jci.insight.93487.
McKinley, E. T., Sui, Y., Al-Kofahi, Y., Millis, B. A., Tyska, M. J., Roland, J. T., Santamaria-Pang, A., Ohland, C. L., Jobin, C., Franklin, J. L., Lau, K. S., Gerdes, M. J., & Coffey, R. J. (2017). Optimized multiplex immunofluorescence single-cell analysis reveals tuft cell heterogeneity. JCI insight, 2(11), . https://doi.org/10.1172/jci.insight.93487
McKinley, Eliot T, et al. "Optimized multiplex immunofluorescence single-cell analysis reveals tuft cell heterogeneity." JCI insight vol. 2,11 (2017). doi: https://doi.org/10.1172/jci.insight.93487
McKinley ET, Sui Y, Al-Kofahi Y, Millis BA, Tyska MJ, Roland JT, Santamaria-Pang A, Ohland CL, Jobin C, Franklin JL, Lau KS, Gerdes MJ, Coffey RJ. Optimized multiplex immunofluorescence single-cell analysis reveals tuft cell heterogeneity. JCI Insight. 2017 Jun 02;2(11). doi: 10.1172/jci.insight.93487. eCollection 2017 Jun 02. PMID: 28570279; PMCID: PMC5453701.
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JCI Insight. 2017 Jun 02;2(11). doi: 10.1172/jci.insight.93487. eCollection 2017 Jun 02.

Optimized multiplex immunofluorescence single-cell analysis reveals tuft cell heterogeneity.

JCI insight

Eliot T McKinley, Yunxia Sui, Yousef Al-Kofahi, Bryan A Millis, Matthew J Tyska, Joseph T Roland, Alberto Santamaria-Pang, Christina L Ohland, Christian Jobin, Jeffrey L Franklin, Ken S Lau, Michael J Gerdes, Robert J Coffey

Affiliations

  1. Epithelial Biology Center and.
  2. Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
  3. General Electric Global Research Center, Niskayuna, New York, USA.
  4. Department of Cell and Developmental Biology.
  5. Cell Imaging Shared Resource, and.
  6. Department of Surgery, Vanderbilt University School of Medicine, Nashville, Tennessee, USA.
  7. Department of Medicine.
  8. Department of Infectious Diseases and Pathology, and.
  9. Department of Anatomy and Cell Physiology, University of Florida, Gainesville, Florida, USA.
  10. Veterans Affairs Medical Center, Tennessee Valley Healthcare System, Nashville, Tennessee, USA.

PMID: 28570279 PMCID: PMC5453701 DOI: 10.1172/jci.insight.93487

Abstract

Intestinal tuft cells are a rare, poorly understood cell type recently shown to be a critical mediator of type 2 immune response to helminth infection. Here, we present advances in segmentation algorithms and analytical tools for multiplex immunofluorescence (MxIF), a platform that enables iterative staining of over 60 antibodies on a single tissue section. These refinements have enabled a comprehensive analysis of tuft cell number, distribution, and protein expression profiles as a function of anatomical location and physiological perturbations. Based solely on DCLK1 immunoreactivity, tuft cell numbers were similar throughout the mouse small intestine and colon. However, multiple subsets of tuft cells were uncovered when protein coexpression signatures were examined, including two new intestinal tuft cell markers, Hopx and EGFR phosphotyrosine 1068. Furthermore, we identified dynamic changes in tuft cell number, composition, and protein expression associated with fasting and refeeding and after introduction of microbiota to germ-free mice. These studies provide a foundational framework for future studies of intestinal tuft cell regulation and demonstrate the utility of our improved MxIF computational methods and workflow for understanding cellular heterogeneity in complex tissues in normal and disease states.

Keywords: Gastroenterology

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