Case Snapshot

The Challenge

A life-science research team needed to resolve nanoscale morphological changes in individual red blood cells associated with aging and oxidative stress, correlating them with optical observations.

The Breakthrough

Correlative optical and AFM imaging with Park NX12 enabled nanoscale morphological analysis of individual red blood cells under controlled, low-force measurement conditions.

The Outcome

The team revealed distinct aging patterns in normal and favism erythrocytes, including morphological alterations, enhanced vesicle formation, and nanomechanical changes.

Why It Matters

The work demonstrates how correlative optical-AFM analysis can reveal nanoscale cellular changes beyond conventional optical observations, supporting deeper investigation of red blood cell aging and disease-related alterations.

Revealing Nanoscale Structural Changes in Red Blood Cells

CNR researchers used Park NX12 for correlative optical-AFM analysis, revealing morphological and nanomechanical changes associated with aging and oxidative stress in individual red blood cells.

  • Customer: Institute of Structure of Matter (ISM), National Research Council of Italy (CNR)
  • Research Focus: Aging, Oxidative Stress Markers and Morpho-Mechanical Changes in Red Blood Cells
  • Solution: Park NX12 for Correlative Optical-AFM and Nanomechanical Analysis
  • Key Value: Nanoscale single-cell characterization under controlled, low-force imaging conditions

The Challenge

Red blood cells undergo progressive morphological and structural changes during aging, which are strongly influenced by oxidative stress. This is particularly relevant in favism, in which G6PD gene deficiency reduces the cells’ ability to respond to oxidative stress.

While the biochemical effects of G6PD deficiency are well established, its nanostructural and nanomechanical consequences at the single-cell level remain less understood. The research team needed to correlate nanoscale membrane morphology with local mechanical properties on the same individual cells to understand how favism erythrocytes evolve during aging.

“Understanding aging and oxidative stress affect individual RBCs requires connecting optical observations with nanoscale structural information. Correlative microscopy provides a powerful way to examine these changes at the single-cell level.”

— Giovanni Longo, Institute of Structure of Matter, CNR, Italy

The Solution

Park NX12 provided the AFM platform for correlating optical observations with nanoscale morphological and nanomechanical characterization of individual red blood cells.

  • Correlative Optical-AFM Imaging: Integrated with an inverted optical microscope to locate individual erythrocytes and perform AFM imaging of the same area.
  • Controlled Low-Force Imaging: Acquired AFM morphological images in non-contact or in soft contact with tip-sample interaction below 2 nN.
  • Nanoscale Morphological Analysis: Characterized cell morphology and membrane roughness across different stages of erythrocyte aging.
  • Nanomechanical Characterization: Mapped Young’s modulus on the same cell areas used for roughness analysis to correlate local mechanical and morphometric properties.

The Results

Correlative optical, fluorescence, and AFM measurements revealed distinct aging behaviors between normal and favism red blood cells. Favism erythrocytes showed a higher occurrence of abnormal cell shapes and increased vesicle formation as aging progressed. AFM analysis of membrane morphology, roughness, and nanomechanical properties further revealed a transition at intermediate-to-late aging stages, associated with reduced and increasingly heterogeneous cytoskeletal support. Together with fluorescence observations, these results provided new insight into the coupling between structural and hemoglobin-related degradation processes underlying accelerated cell senescence in favism.

“Park NX12 allowed us to examine the same red blood cells identified by optical microscopy at the nanoscale, providing detailed morphological and nanomechanical information on their structural changes during oxidative aging.”

— Marco Girasole, Institute of Structure of Matter, CNR, Italy

Key Outcomes

  • Revealed distinct aging patterns in normal and favism red blood cells
  • Identified increased morphological alterations and vesicle formation in aging favism erythrocytes
  • Characterized changes in membrane roughness and nanomechanical properties during oxidative aging
  • Combined nanoscale structural analysis with fluorescence observations of hemoglobin-related alterations

Related Publication

This capability is further demonstrated through peer-reviewed research from the Institute of Structure of Matter, CNR, on oxidative aging and morpho-mechanical changes in favism erythrocytes, published in International Journal of Molecular Sciences (DOI: 10.3390/ijms27031132) and on procedures used to perform such analyses, published in Methods and Protocols (DOI: 10.3390/mps8040086).