Three-Dimensional Reconstruction of Toxoplasma–Neuron Interactions In Situ

Carla M. Cabral, Emily F. Merritt, Anita A. Koshy

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

How tissue and cellular architecture affects host cell–microbe interactions in vivo remains poorly defined because imaging these interactions in complex tissue is difficult and standard in vitro cultures do not mimic whole organ architecture. Here we describe a method that combines new tissue clearing techniques, high-resolution imaging, and three-dimensional reconstruction to overcome these barriers and allow in situ imaging of host cell–microbe interactions in complex tissue. We use the interactions between neurons and Toxoplasma gondii, a ubiquitous, protozoan parasite that establish a lifelong central nervous system (CNS) infection in mice and humans, as a model for this technique. This method aims to provide an easy, reproducible way to visualize the complex relationship between host cells and intracellular pathogens within a whole organ.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages283-295
Number of pages13
DOIs
StatePublished - Jan 1 2020

Publication series

NameMethods in Molecular Biology
Volume2071
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • 3-D reconstruction
  • Confocal microscopy
  • Host-microbe interactions
  • Tissue clearing
  • Toxoplasma

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Cabral, C. M., Merritt, E. F., & Koshy, A. A. (2020). Three-Dimensional Reconstruction of Toxoplasma–Neuron Interactions In Situ. In Methods in Molecular Biology (pp. 283-295). (Methods in Molecular Biology; Vol. 2071). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9857-9_16