Label-Free Digital Holotomography Reveals Ibuprofen-Induced Morphological Changes to Red Blood Cells

Talia Bergaglio, Shayon Bhattacharya, Damien Thompson, Peter Niraj Nirmalraj

Research output: Contribution to journalArticlepeer-review

Abstract

Understanding the dose-dependent effect of over-the-counter drugs on red blood cells (RBCs) is crucial for hematology and digital pathology. Yet, it is challenging to continuously record the real-time, drug-induced shape changes of RBCs in a label-free manner. Here, we demonstrate digital holotomography (DHTM)-enabled real-time, label-free concentration-dependent and time-dependent monitoring of ibuprofen on RBCs from a healthy donor. The RBCs are segmented based on three-dimensional (3D) and four-dimensional (4D) refractive index tomograms, and their morphological and chemical parameters are retrieved with their shapes classified using machine learning. We directly observed the formation and motion of spicules on the RBC membrane when aqueous solutions of ibuprofen were drop-cast on wet blood, creating rough-membraned echinocyte forms. At low concentrations of 0.25-0.50 mM, the ibuprofen-induced morphological change was transient, but at high concentrations (1-3 mM) the spiculated RBC remained over a period of up to 1.5 h. Molecular simulations confirmed that aggregates of ibuprofen molecules at high concentrations significantly disrupted the RBC membrane structural integrity and lipid order but produced negligible effect at low ibuprofen concentrations. Control experiments on the effect of urea, hydrogen peroxide, and aqueous solutions on RBCs showed zero spicule formation. Our work clarifies the dose-dependent chemical effects on RBCs using label-free microscopes that can be deployed for the rapid detection of overdosage of over-the-counter and prescribed drugs.

Original languageEnglish
Pages (from-to)241-255
Number of pages15
JournalACS Nanoscience Au
Volume3
Issue number3
DOIs
Publication statusPublished - 21 Jun 2023

Keywords

  • digital holotomography
  • ibuprofen
  • machine learning
  • molecular dynamics simulations
  • red blood cells

Fingerprint

Dive into the research topics of 'Label-Free Digital Holotomography Reveals Ibuprofen-Induced Morphological Changes to Red Blood Cells'. Together they form a unique fingerprint.

Cite this