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HoloBio: A holographic microscopy tool for quantitative biological analysis.

Waira Mona1, Maria J Gil-Herrera1, Emanuel Mazo1

  • 1Applied Optics & Electronic Instrumentation Laboratory, School of Applied Science and Engineering, Universidad EAFIT, Medellín, Colombia.

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|June 22, 2026
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Summary

HoloBio software provides accessible, open-source tools for holographic microscopy analysis. This Python-based graphical user interface enables label-free, quantitative imaging of biological specimens for researchers.

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Area of Science:

  • Biophysics and quantitative imaging
  • Cell biology and morphology analysis
  • Biomedical optics and photonics

Background:

  • Holographic imaging offers label-free, quantitative data for biological specimens.
  • Widespread adoption is hindered by a lack of accessible and standardized analysis software.
  • Current tools often require significant coding expertise, limiting accessibility for biologists.

Purpose of the Study:

  • To develop an open-source, user-friendly software for holographic microscopy analysis.
  • To provide accessible tools for quantitative analysis of biological samples using holography.
  • To support diverse holographic imaging setups and analysis tasks.

Main Methods:

  • Developed HoloBio, an open-source, Python-based graphical user interface (GUI).
  • Implemented Real-Time and Offline processing modes for live and post-processing of holograms.
  • Ensured compatibility with various lens-based/lensless and off-axis/in-line holographic configurations.

Main Results:

  • HoloBio offers tools for cell tracking, phase profiling, thickness estimation, and morphological analysis.
  • Includes functionalities for cell counting and object area quantification.
  • Provides a reproducible, high-throughput analysis environment accessible to non-coders.

Conclusions:

  • HoloBio addresses the need for accessible holographic microscopy analysis software.
  • Facilitates quantitative, label-free imaging in biology, biophotonics, and biomedical imaging.
  • Empowers researchers without coding expertise to leverage holographic imaging techniques.