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Imaging semi-automatico di fluorescenza tessuto-specifica in embrioni di zebrafish
Imaging semi-automatico di fluorescenza tessuto-specifica in embrioni di zebrafish
JoVE Journal
Biology
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JoVE Journal Biology
Semi-automated Imaging of Tissue-specific Fluorescence in Zebrafish Embryos

Imaging semi-automatico di fluorescenza tessuto-specifica in embrioni di zebrafish

Full Text
10,163 Views
07:06 min
May 17, 2014

DOI: 10.3791/51533-v

Shannon N. Romano1, Daniel A. Gorelick1

1Department of Pharmacology and Toxicology,University of Alabama at Birmingham

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This protocol outlines a method for semi-automated imaging of tissue-specific fluorescence in zebrafish embryos, focusing on estrogen responsive cells. The procedure utilizes fluorescent microscopy to visualize the activity of estrogen receptors in live zebrafish larvae.

Key Study Components

Area of Science

  • Neuroscience
  • Developmental Biology
  • Fluorescent Imaging

Background

  • Estrogen receptors play a critical role in various biological processes.
  • Zebrafish serve as a model organism for studying developmental biology.
  • Fluorescent microscopy allows for the visualization of specific cellular activities.
  • Automated imaging techniques enhance the efficiency of data collection.

Purpose of Study

  • To develop a protocol for detecting estrogen responsive cells in zebrafish.
  • To improve the accuracy and efficiency of imaging techniques in live embryos.
  • To contribute to the understanding of estrogen receptor activity in development.

Methods Used

  • Breeding of X-E-R-E-G-F-P adult zebrafish to generate embryos.
  • Placement of embryos in individual wells of a 96 well plate.
  • Addition of estrogens to the wells containing embryos.
  • Scanning of the plate for fluorescence using an epi fluorescent microscope with a motorized stage.

Main Results

  • Successful detection of estrogen responsive cells in live zebrafish larvae.
  • Visualization of tissue-specific activity of estrogen receptors.
  • Demonstration of the effectiveness of the semi-automated imaging protocol.
  • Contribution to the field of developmental biology through enhanced imaging techniques.

Conclusions

  • The protocol provides a reliable method for studying estrogen receptor activity.
  • Automated imaging can significantly streamline research processes.
  • Findings may have implications for understanding hormonal influences on development.

Frequently Asked Questions

What is the main goal of this protocol?
The main goal is to automatically detect estrogen responsive cells in live zebrafish larvae.
How are the embryos prepared for imaging?
Embryos are placed in individual wells of a 96 well plate and treated with estrogens.
What type of microscopy is used in this study?
Epi fluorescent microscopy with a motorized stage is used for imaging.
Why are zebrafish used as a model organism?
Zebrafish are transparent during early development, allowing for easy observation of cellular processes.
What are the implications of this research?
This research may enhance the understanding of hormonal influences on development and improve imaging techniques.

Qui descritto è un protocollo per l'imaging semi-automatica di fluorescenza tessuto-specifica in embrioni di zebrafish.

L'obiettivo generale di questa procedura è quello di rilevare automaticamente le cellule sensibili agli estrogeni nelle larve di zebrafish vive. Ciò si ottiene allevando prima cinque pesci adulti X-E-R-E-G-F-P per generare embrioni di pesce zebra reporter di estrogeni. Successivamente, gli embrioni vengono posti in pozzetti individuali di una piastra a 96 pozzetti.

Quindi gli estrogeni vengono aggiunti alla piastra degli embrioni. Infine, la lastra viene scansionata per la fluorescenza utilizzando un microscopio epifluorescente con tavolino motorizzato. In definitiva, la microscopia fluorescente viene utilizzata per mostrare l'attività tissutale specifica dei recettori degli estrogeni.

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