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Biology
在斑马鱼发育的细胞追踪使用Photoconvertible蛋白
在斑马鱼发育的细胞追踪使用Photoconvertible蛋白
JoVE Journal
Biology
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JoVE Journal Biology
Cell Tracking Using Photoconvertible Proteins During Zebrafish Development

在斑马鱼发育的细胞追踪使用Photoconvertible蛋白

Full Text
11,984 Views
07:19 min
September 28, 2012

DOI: 10.3791/4350-v

Verónica A. Lombardo1, Anje Sporbert1, Salim Abdelilah-Seyfried1

1Max Delbrück Center for Molecular Medicine

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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 article presents a method for tracking photo switched cells in living zebrafish embryos using photoconvertible fluorescent proteins. The technique allows for the monitoring of cell biological events during development through confocal microscopy.

Key Study Components

Area of Science

  • Neuroscience
  • Cell Biology
  • Developmental Biology

Background

  • Photoconvertible fluorescent proteins (PCFPs) enable visualization of specific cells.
  • Tracking cellular events in live organisms is crucial for understanding development.
  • Zebrafish embryos are a model organism for studying vertebrate development.
  • Confocal microscopy provides high-resolution imaging of biological samples.

Purpose of Study

  • To develop a method for tracking photo switched cells in zebrafish embryos.
  • To monitor the stability of photoconverted proteins during development.
  • To enhance understanding of developmental processes in live vertebrates.

Methods Used

  • Embedding zebrafish embryos in low melting temperature agarose.
  • Expressing tissue-specific photoconvertible fluorescent proteins.
  • Photo converting proteins in the region of interest.
  • Maintaining embryos in egg water at 28.5 degrees Celsius for development.

Main Results

  • Photo converted cells can be detected at later developmental stages.
  • Stable presence of photoconverted proteins allows for precise tracking.
  • Confocal microscopy effectively visualizes the photo switched cells.
  • The methodology can be applied to various developmental studies.

Conclusions

  • The presented method is effective for tracking cellular events in live zebrafish embryos.
  • It provides insights into developmental processes and cell biology.
  • This approach can be utilized in future studies of vertebrate development.

Frequently Asked Questions

What are photoconvertible fluorescent proteins?
Photoconvertible fluorescent proteins are proteins that change their fluorescence properties upon exposure to specific light wavelengths, allowing for the tracking of cells.
Why are zebrafish embryos used in this study?
Zebrafish embryos are transparent and develop rapidly, making them ideal for observing developmental processes in real-time.
What is confocal microscopy?
Confocal microscopy is an imaging technique that provides high-resolution images by using a focused laser beam to scan samples.
How does the photo switching process work?
The photo switching process involves exposing the fluorescent protein to specific light, causing it to change its fluorescence characteristics and allowing for tracking.
What are the potential applications of this methodology?
This methodology can be applied to various studies in developmental biology, cell biology, and neuroscience to track cellular events.
What temperature is optimal for maintaining zebrafish embryos?
The optimal temperature for maintaining zebrafish embryos during development is 28.5 degrees Celsius.

在这里,我们提出了一种在活斑马鱼胚胎中光转化荧光蛋白 (PCFPs) 进行光激活开关的方法,并在发育过程中的特定时间点进一步跟踪光转化蛋白。该方法允许监测活体脊椎动物生物体中不同发育过程背后的细胞生物事件。

该程序的总体目标是追踪活斑马鱼胚胎中的光开关细胞。这是通过首先包埋胚胎,在低熔点温度下表达组织特异性光、可转换荧光蛋白来实现的。接下来,对感兴趣区域内的蛋白质进行完全光转化。

然后小心地将胚胎从 aros 中取出,并在 28.5 摄氏度的鸡蛋水中保持直到后期。最后,光转化的胚胎再次嵌入低熔融温度的 aros 中。最终,可以获得结果表明,由于通过共聚焦显微镜稳定存在光转化蛋白,可以在后期精确检测到光转化细胞。

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细胞追踪 光转化蛋白 斑马鱼发育 胚胎发生 基因编码荧光蛋白 动态形态发生过程 光激活蛋白 光转化荧光蛋白 (PCFPs) Kaede KikGR Dendra EosFP 绿到红移 紫外线或蓝光曝光 细胞命运追踪

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