Non-invasive Imaging of the Innate Immune Response in a Zebrafish Larval Model of <em>Streptococcus iniae</em> Infection

Elizabeth A. Harvie; Anna Huttenlocher

doi:10.3791/52788

Imagerie non invasive de la réponse immunitaire innée dans un modèle de poisson zèbre larves de

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Immunology and Infection

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JoVE Journal Immunology and Infection

Non-invasive Imaging of the Innate Immune Response in a Zebrafish Larval Model of Streptococcus iniae Infection

Imagerie non invasive de la réponse immunitaire innée dans un modèle de poisson zèbre larves de Streptococcus iniae Infection

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11:16 min

April 21, 2015

DOI: 10.3791/52788-v

Elizabeth A. Harvie^1,2, Anna Huttenlocher^2,3

¹Microbiology Doctoral Training Program,University of Wisconsin-Madison, ²Department of Medical Microbiology and Immunology,University of Wisconsin-Madison, ³Department of Pediatrics,University of Wisconsin-Madison

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Overview

This article presents a protocol for microinjecting zebrafish larvae with the bacterial pathogen Streptococcus IEA to study the host immune response. The procedure includes live imaging techniques to visualize the infection in the otic vesicle.

Key Study Components

Area of Science

Neuroscience
Microbiology
Immunology

Background

Zebrafish are a valuable model for studying immune responses.
Microinjection techniques allow for precise delivery of pathogens.
Live imaging provides insights into host-pathogen interactions.
Confocal microscopy enhances visualization of immune cells.

Purpose of Study

To develop a method for inducing localized bacterial infections in zebrafish.
To visualize and track the immune response to bacterial infection.
To utilize photo labeling for enhanced imaging of immune cells.

Methods Used

Microinjection of zebrafish larvae with Streptococcus IEA.
Use of agarose molds for positioning larvae during injection.
Photo labeling of host immune cells for tracking.
Imaging of infections using confocal microscopy.

Main Results

Successful microinjection into the otic vesicle of zebrafish larvae.
Visualization of the immune response to the bacterial infection.
Effective tracking of labeled immune cells during imaging.
Demonstration of the utility of zebrafish for studying host-pathogen interactions.

Conclusions

The protocol enables detailed study of immune responses in zebrafish.
Live imaging techniques provide valuable insights into infection dynamics.
This method can be applied to other pathogens for broader research.

Frequently Asked Questions

What is the significance of using zebrafish in this study?

Zebrafish are transparent during early development, allowing for direct observation of immune responses in real-time.

How does microinjection work?

Microinjection involves using a fine needle to deliver a bacterial suspension directly into the otic vesicle of the zebrafish larvae.

What imaging technique is used in this study?

Confocal microscopy is used to visualize the infection and track immune cells.

What are the advantages of photo labeling immune cells?

Photo labeling allows for easy tracking of immune cells during imaging, enhancing the understanding of their behavior during infection.

Can this method be applied to other pathogens?

Yes, the protocol can be adapted for studying various bacterial and viral pathogens in zebrafish.

Ici, nous présentons un protocole pour la génération et l’imagerie d’une infection bactérienne localisée dans la vésicule otique du poisson zèbre.

L’objectif global de cette procédure est de micro-injecter à la larve de poisson-zèbre un pathogène bactérien streptocoque IEA pour une imagerie non invasive en direct de la réponse immunitaire de l’hôte. Ceci est accompli en chargeant d’abord l’aiguille de micro-injection avec une suspension bactérienne et en alignant les larves de poisson-zèbre anesthésiées sur le moule d’injection d’agarose. Ensuite, la suspension bactérienne est micro-injectée dans la vésicule otique des larves.

Ensuite, les larves injectées sont montées dans de l’agarose à bas point de fusion. Enfin, les cellules immunitaires de l’hôte Gus sont photomarquées pour faciliter le suivi et l’infection est imagée à l’aide d’un microscope confocal. En fin de compte, la micro-injection de streptocoque IEA dans la vésicule otique et le photomarquage et la microscopie confocale qui s’ensuivent sont utilisés pour étudier la réponse immunitaire de l’hôte des larves de poisson-zèbre.

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