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Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...
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An Automated Microscopic Scoring Method for the &#947;-H2AX Foci Assay in Human Peripheral Blood Lymphocytes
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Foci-Xpress: Automated and Fast Nuclear Foci Counting Tool.

Jae-I Moon1,2, Woo-Jin Kim1,2, Ki-Tae Kim1,2

  • 1Department of Molecular Genetics and Dental Pharmacology, School of Dentistry and Dental Research Institute, Dental Multi-Omics Center, Seoul National University, Seoul 08826, Republic of Korea.

International Journal of Molecular Sciences
|October 14, 2023
PubMed
Summary
This summary is machine-generated.

Automated software Foci-Xpress precisely quantifies subcellular foci in the cell nucleus. This open-source tool offers faster, unbiased counting compared to manual methods, aiding cellular dynamics research.

Keywords:
DNA damageautomated screening methodfoci quantitative analysisimage processingnuclear foci

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Bioimaging

Background:

  • Subcellular foci in the nucleus perform specific molecular functions.
  • Accurate quantification of these foci is crucial for understanding cellular dynamics and signaling.
  • Manual foci counting is laborious and prone to errors.

Purpose of the Study:

  • To develop an automated image analysis method for precise quantification of nuclear foci.
  • To introduce Foci-Xpress, an open-source software for automatic foci counting.
  • To compare Foci-Xpress with conventional methods regarding efficiency, speed, accuracy, and convenience.

Main Methods:

  • Development of an open-source software (Foci-Xpress) for automated foci counting.
  • Image analysis incorporating adjustable brightness thresholds for foci detection.
  • Comparative analysis of Foci-Xpress against conventional foci counting techniques using induced models.

Main Results:

  • Foci-Xpress demonstrated significantly higher speed compared to conventional methods.
  • The accuracy of Foci-Xpress was comparable to traditional approaches.
  • Automation by Foci-Xpress eliminated the need for specialized analysis equipment and researcher bias.

Conclusions:

  • Foci-Xpress provides a fast, accurate, and convenient automated solution for quantifying nuclear foci.
  • The software enhances experimental throughput and facilitates robust statistical analysis.
  • Foci-Xpress is an accessible tool for cell biologists, reducing manual errors and variations.