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Related Experiment Video

Updated: May 26, 2026

Single Cell Fate Mapping in Zebrafish
07:53

Single Cell Fate Mapping in Zebrafish

Published on: October 5, 2011

Single cell fate mapping in zebrafish.

Vikram Kohli1, Kira Rehn, Saulius Sumanas

  • 1Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, USA.

Journal of Visualized Experiments : Jove
|December 14, 2011
PubMed
Summary
This summary is machine-generated.

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This study introduces a new method for cell tracing in embryos using near-infrared laser pulses to activate caged fluorescein-dextran. This technique allows for precise, deep-tissue single-cell labeling and subsequent detection for developmental biology research.

Area of Science:

  • Developmental Biology
  • Cell Biology
  • Microscopy

Background:

  • Accurate cell fate mapping and lineage tracing are crucial for understanding embryonic development, particularly for hematopoietic, lymphatic, and blood vessel lineages.
  • Photoactivatable proteins and caged fluorescein-dextran are used for cell tracing, but traditional UV-based uncaging methods lack spatial resolution and can limit long-term tracking.
  • Existing methods using UV excitation for uncaging fluorescein-dextran have limitations in spatial resolution, hindering precise single-cell labeling deep within biological tissues.

Purpose of the Study:

  • To develop and present a novel protocol for precise single-cell fate mapping and lineage tracing in developing embryos.
  • To overcome the limitations of UV-based photoactivation by employing near-infrared laser pulses for enhanced spatial resolution and depth penetration.
  • To establish a versatile method for activating and detecting single labeled cells applicable to various model systems.

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Isolation and Characterization of Single Cells from Zebrafish Embryos
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Isolation and Characterization of Single Cells from Zebrafish Embryos

Published on: March 12, 2016

Lineage Labeling of Zebrafish Cells with Laser Uncagable Fluorescein Dextran
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Lineage Labeling of Zebrafish Cells with Laser Uncagable Fluorescein Dextran

Published on: April 28, 2011

Related Experiment Videos

Last Updated: May 26, 2026

Single Cell Fate Mapping in Zebrafish
07:53

Single Cell Fate Mapping in Zebrafish

Published on: October 5, 2011

Isolation and Characterization of Single Cells from Zebrafish Embryos
09:25

Isolation and Characterization of Single Cells from Zebrafish Embryos

Published on: March 12, 2016

Lineage Labeling of Zebrafish Cells with Laser Uncagable Fluorescein Dextran
07:35

Lineage Labeling of Zebrafish Cells with Laser Uncagable Fluorescein Dextran

Published on: April 28, 2011

Main Methods:

  • Injection of caged fluorescein-dextran into single cells within developing embryos.
  • Photoactivation of caged fluorescein-dextran using near-infrared laser pulses from a titanium sapphire femtosecond laser, enabling two-photon absorption.
  • Utilizing a two-photon confocal microscope (LSM 510 META NLO) for precise laser focusing deep within the embryo.
  • Development of a simple immunohistochemistry protocol for rapid visualization of the activated single cell.

Main Results:

  • Demonstrated successful photoactivation of caged fluorescein-dextran in single cells deep within embryos using near-infrared laser pulses.
  • Achieved high spatial resolution, uncaging only at the laser's focal point without affecting surrounding cells.
  • Successfully visualized the activated cells using the described immunohistochemistry protocol, enabling detection of labeled cells.

Conclusions:

  • The reported protocol offers a significant advancement in single-cell fate mapping and lineage tracing by enabling precise, deep-tissue labeling.
  • The use of near-infrared laser pulses and two-photon absorption overcomes the spatial resolution limitations of traditional UV-based methods.
  • This versatile activation and detection strategy is broadly applicable to diverse embryonic model systems for detailed developmental studies.