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DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
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Fluorescent Photo-conversion: A second chance to label unique cells.

Adam J Mellott1, Heather E Shinogle2, David S Moore2

  • 1Bioengineering Graduate Program, University of Kansas, Lawrence, Kansas 66045.

Cellular and Molecular Bioengineering
|April 28, 2015
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Summary

Researchers can now isolate unique cells from treated populations using Dendra2 photo-convertible proteins. This method enables precise identification and study of individual cells, advancing tissue engineering research.

Keywords:
Cell trackingDendra2Human Wharton's jelly cellsNucleofectionPhoto-convertible reporter gene

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Area of Science:

  • Cell Biology
  • Biotechnology
  • Tissue Engineering

Background:

  • Cellular heterogeneity complicates tissue engineering studies.
  • Identifying unique cellular responses to treatment is crucial for understanding treatment efficacy.
  • Existing methods lack precision in isolating specific cell populations within a larger group.

Purpose of the Study:

  • To demonstrate the use of photo-convertible reporter proteins for selective cell labeling.
  • To establish a method for identifying and isolating unique human Wharton's Jelly cells (hWJCs) after treatment.
  • To enable more detailed analysis of individual cell behavior within a treated population.

Main Methods:

  • Transfection of hWJCs with Dendra2 reporter protein.
  • Selective photo-conversion of Dendra2 from green to red fluorescence in specific cells.
  • Utilizing the green-to-red fluorescence shift for precise cell identification.

Main Results:

  • Consistent and robust photo-conversion of Dendra2 was achieved in selected hWJCs.
  • Arbitrarily selected cells were precisely identified and labeled via photo-conversion.
  • The method successfully distinguished unique cells within the treated population.

Conclusions:

  • Photo-convertible proteins like Dendra2 offer a powerful tool for selective cell labeling and identification.
  • This technique allows for the isolation and further study of unique cells within a heterogeneous population.
  • The method enhances the ability to gain meaningful insights into treatment effects on individual cells in tissue engineering.