Optimized immunofluorescence staining protocol for imaging germinal centers in secondary lymphoid tissues of vaccinated mice

STAR Protoc. 2021 Jun 18;2(3):100499. doi: 10.1016/j.xpro.2021.100499. eCollection 2021 Sep 17.

Abstract

Location of immune cells that form the germinal center reaction within secondary lymphoid tissues can be characterized using confocal microscopy. Here, we present an optimized immunofluorescence staining protocol to image germinal center structures in fixed/frozen spleen sections from ChAdOx1 nCoV-19 immunized mice. This protocol can be adapted to identify other cell types within secondary lymphoid tissues. For complete information on the generation and use of this protocol to examine immune responses to the COVID vaccine ChAdOx1 nCoV-19, please refer to Silva-Cayetano et al. (2020).

Keywords: Antibody; Cell Biology; Immunology; Microbiology; Microscopy; Model Organisms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • COVID-19 / diagnostic imaging
  • COVID-19 / immunology
  • COVID-19 / prevention & control*
  • COVID-19 / virology
  • ChAdOx1 nCoV-19 / administration & dosage*
  • Fluorescent Antibody Technique / methods
  • Fluorescent Antibody Technique / standards*
  • Germinal Center / drug effects*
  • Germinal Center / immunology
  • Germinal Center / pathology
  • Germinal Center / virology
  • Immunization, Secondary / methods*
  • Immunogenicity, Vaccine
  • Male
  • Mice
  • SARS-CoV-2 / immunology*
  • Spleen / drug effects*
  • Spleen / immunology
  • Spleen / pathology
  • Spleen / virology

Substances

  • ChAdOx1 nCoV-19