Validation of an organ mapping antibody panel for cyclical immunofluorescence microscopy on normal human kidneys

Author:

Brewer Maya1ORCID,Migas Lukasz G.2,Clouthier Kelly A.1,Allen Jamie L.34,Anderson David M.35,Pingry Ellie34,Farrow Melissa35,Quardokus Ellen M.6,Spraggins Jeffrey M.34578,Van de Plas Raf235,de Caestecker Mark P.14ORCID

Affiliation:

1. Division of Nephrology, Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee, United States

2. Delft Center for Systems and Control, Delft University of Technology, Delft, The Netherlands

3. Mass Spectrometry Research Center, Vanderbilt University School of Medicine, Nashville, Tennessee, United States

4. Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, Tennessee, United States

5. Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee, United States

6. Department of Intelligent Systems Engineering, Indiana University, Bloomington, Indiana, United States

7. Department of Chemistry, Vanderbilt University School of Medicine, Nashville, Tennessee, United States

8. Department of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee, United States

Abstract

We describe here validation criteria used to define on organ mapping panel of antibodies that can be used to define 18 cell types and five extracellular matrix compartments using cyclical immunofluorescence (CyCIF) microscopy. As CyCIF does not require specialized instrumentation, and image registration required to assemble CyCIF images can be performed by any laboratory without specialized computational skills, this technology is accessible to any laboratory with access to a fluorescence microscope and digital scanner.

Funder

HHS | NIH | National Institute of Diabetes and Digestive and Kidney Diseases

HHS | NIH | OSC | Common Fund

Publisher

American Physiological Society

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