Long-term Live Imaging Device for Improved Experimental Manipulation of Zebrafish Larvae

Kayla Huemer; Jayne M. Squirrell; Robert Swader; Kirsten Pelkey; Danny C. LeBert; Anna Huttenlocher; Kevin W. Eliceiri

doi:10.3791/56340

Dispositivo de proyección de imagen de vivo a largo plazo para mejor manipulación Experimental de...

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Bioengineering

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JoVE Journal Bioengineering

Long-term Live Imaging Device for Improved Experimental Manipulation of Zebrafish Larvae

Dispositivo de proyección de imagen de vivo a largo plazo para mejor manipulación Experimental de larvas de pez cebra

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14:05 min

October 27, 2017

DOI: 10.3791/56340-v

Kayla Huemer*^1,2, Jayne M. Squirrell*³, Robert Swader², Kirsten Pelkey², Danny C. LeBert⁴, Anna Huttenlocher^5,6, Kevin W. Eliceiri^1,2,3

¹Department of Biomedical Engineering,University of Wisconsin-Madison, ²Morgridge Institute for Research,University of Wisconsin-Madison, ³Laboratory for Optical and Computational Instrumentation,University of Wisconsin-Madison, ⁴Cellular and Molecular Pathology Graduate Program,University of Wisconsin-Madison, ⁵Department of Medical Microbiology and Immunology,University of Wisconsin-Madison, ⁶Department of Pediatrics,University of Wisconsin-Madison

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Overview

This manuscript describes the zWEDGI (zebrafish Wounding and Entrapment Device for Growth and Imaging), a device designed to orient and restrain zebrafish larvae for high-resolution imaging of wound healing. This method allows for tail transection and long-term observation of regeneration processes.

Key Study Components

Area of Science

Neuroscience
Biology
Regenerative Medicine

Background

The zWEDGI device facilitates the study of wound healing in live zebrafish larvae.
It allows for simultaneous wounding and imaging without hindering tail regrowth.
The device's compartments can be modified for different experimental protocols.
This approach addresses key questions regarding collagen reorganization and cellular responses post-injury.

Purpose of Study

To create a platform for microscopic analysis of wound healing.
To investigate the contributions of various biological factors in tissue regrowth.
To develop a versatile device for different larval stages and experimental needs.

Methods Used

Modeling the PDMS component of the device according to specified geometries.
Using a photopolymer 3-D printer to create molds.
Cleaning the molds with denatured alcohol and compressed air.
Implementing the device for live imaging of zebrafish larvae.

Main Results

The zWEDGI effectively supports long-term imaging of wound healing.
Tail regrowth was not impeded by the device during experiments.
High-resolution images provided insights into the healing process.
The device's design allows for modifications to suit various experimental setups.

Conclusions

The zWEDGI is a valuable tool for studying wound healing in zebrafish larvae.
This device enables researchers to explore critical biological processes in real-time.
Future studies can utilize this platform to further understand tissue regeneration mechanisms.

Frequently Asked Questions

What is the zWEDGI device?

The zWEDGI is a compartmentalized device designed for imaging and analyzing wound healing in zebrafish larvae.

How does the zWEDGI facilitate imaging?

It allows for simultaneous wounding and imaging without hindering the natural regrowth of the tail.

What are the main advantages of using the zWEDGI?

The device supports long-term imaging and can be modified for various experimental protocols.

What biological processes can be studied with this device?

It can be used to investigate collagen reorganization and cellular responses during tissue regeneration.

What materials are used in the zWEDGI?

The device is primarily made from PDMS, which is modeled and fabricated using 3-D printing techniques.

Can the zWEDGI be adapted for different larval stages?

Yes, the multiple compartments can be modified to accommodate different stages of zebrafish larvae.

Este manuscrito describe la zWEDGI (pez cebra herir y el dispositivo de captura para el crecimiento y la proyección de imagen), que es un dispositivo compartimentado diseñado para orientar y restringir las larvas de pez cebra. El diseño permite el corte transversal de la cola y a largo plazo colección de imágenes de alta resolución microscopia fluorescente de la cicatrización de heridas y regeneración.

El objetivo general de la fabricación de este dispositivo de imagen es crear una plataforma modificable para el análisis microscópico de la cicatrización de heridas en larvas vivas de pez cebra. Este método puede ayudar a responder preguntas clave en el campo de la cicatrización de heridas, como cuál es la contribución de la reorganización de las fibras de colágeno y las respuestas celulares al posterior recrecimiento del tejido después de una lesión. La principal ventaja de esta técnica es que las heridas y las imágenes se producen dentro de un solo dispositivo, mientras que el dispositivo no impide el recrecimiento de la cola.

Además, los múltiples compartimentos funcionales del zWEDGI podrían modificarse de forma independiente para adaptarse a diferentes estadios larvarios y a una serie de protocolos experimentales. Para comenzar, modele el componente PDMS del dispositivo con las geometrías y atributos deseados de acuerdo con el protocolo de texto. Después de imprimir los moldes con una impresora 3D de fotopolímero, limpie los moldes con un cepillo fino, alcohol desnaturalizado en una botella de spray y aire comprimido.

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