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A novel AI-coupled flow chamber method quantifying erythrocyte osmotic fragility.

Ipek Seda Fırat1, Özgür Alaçayır1, Till Creutz1,2

  • 1Center of Competence for Bioengineering, University of Applied Sciences Aachen, Medical and Biological Laboratory, Jülich, Germany.

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Summary

A new flow chamber platform with AI analysis automates red blood cell (RBC) osmotic fragility testing. This novel BioExP system accurately measures RBC membrane stability and detects changes induced by inhibitors.

Keywords:
AnemiaArtificial intelligenceBlood bankingErythrocytesOsmotic fragilitySepsis

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Area of Science:

  • Hematology
  • Biophysics
  • Medical Technology

Background:

  • Osmotic fragility (OF) testing traditionally assesses red blood cell (RBC) membrane stability using spectrophotometric, visual, or flow cytometric methods.
  • Existing techniques can be labor-intensive and lack automation for reproducible analysis.
  • There is a need for advanced platforms that offer automated, sensitive, and real-time OF assessment.

Purpose of the Study:

  • To introduce and validate a novel flow chamber-based platform (BioExP) with AI-driven RBC detection for automated OF analysis.
  • To compare the performance of the BioExP platform against classical OF methods.
  • To evaluate the biological sensitivity of the platform using known modulators of RBC membrane integrity.

Main Methods:

  • Development of a flow chamber platform integrated with proprietary imaging software and AI for RBC detection.
  • Comparison of median corpuscular fragility (MCF₅₀) values between the BioExP system and classical methods using samples from healthy donors.
  • Optimization of the protocol, including determining "satiation time" and hemolysis kinetics.
  • Testing biological sensitivity using aquaporin (AQP) inhibition (HgCl₂) and lipopolysaccharide (LPS) to induce membrane changes.

Main Results:

  • The BioExP platform demonstrated agreement with classical OF measurements.
  • Protocol optimization identified optimal conditions for inducing hemolysis.
  • Both HgCl₂ (inhibiting AQP channels) and LPS (increasing membrane fragility) caused significant, reproducible shifts in MCF₅₀ values compared to controls.
  • The platform successfully captured donor-specific variability and detected treatment-induced changes in RBC membrane integrity.

Conclusions:

  • The novel BioExP platform provides an automated, reproducible, and sensitive method for OF analysis.
  • The platform accurately replicates classical OF measurements and detects biological modulations of RBC membrane stability.
  • The study highlights the capability of LPS to compromise RBC membrane integrity independently in plasma-free conditions.
  • BioExP offers advantages such as minimal sample volume requirements and real-time, multi-condition testing capabilities.