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

JoVE Journal
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

先天免疫反应中的斑马鱼幼体型号无创成像链球菌iniae感染

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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.

在这里，我们提出了一种在斑马鱼耳囊中产生和成像局部细菌感染的方案。

该程序的总体目标是用细菌病原体链球菌 IEA 显微注射斑马鱼幼虫，以对宿主免疫反应进行实时无创成像。这是通过首先用细菌悬浮液加载显微注射针并将麻醉的斑马鱼幼虫排列在琼脂糖注射模具上来实现的。接下来，将细菌悬浮液显微注射到幼虫的耳囊中。

然后将注射的幼虫封片在低熔点琼脂糖中。Gus 最后对宿主免疫细胞进行照片标记以便于追踪，并使用共聚焦显微镜对感染进行成像。最终，将链球菌 IEA 显微注射到耳囊中，随后进行照片标记和共聚焦显微镜检查，用于研究斑马鱼幼虫的宿主免疫反应。

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免疫学第98 免疫感染先天性免疫斑马鱼幼体斑马鱼中性粒细胞巨噬细胞链球菌显微注射共焦成像光转化