Analyzing Blood Cells of High-Risk Myelodysplastic Syndrome Patients Using Interferometric Phase Microscopy and...

Area of Science:

• Hematology
• Cell Biology
• Medical Diagnostics

Background:

• Myelodysplastic syndromes (MDS) are serious blood disorders affecting neutrophil morphology and function.
• Early diagnosis of MDS is critical for effective treatment and improved patient prognosis.
• Current diagnostic methods may not fully capture subtle morphological changes in neutrophils.

Purpose of the Study:

• To develop a novel approach for detecting morphological differences in neutrophils between MDS patients and healthy blood bank donors (BBDs).
• To investigate the utility of interferometric phase microscopy (IPM) for MDS diagnosis.

Main Methods:

• Neutrophils were isolated and stained with fluorescent markers (DCF for ROS, Hoechst for DNA) for initial analysis via flow cytometry.
• Fluorescence-activated cell sorting (FACS) was used to isolate distinct neutrophil populations (H and L cells).
• Interferometric phase microscopy (IPM) was employed to image and analyze cell morphology and dry mass without staining.

Main Results:

• Flow cytometry revealed two distinct neutrophil clusters (H and L) in BBDs, but only one (L) in MDS patients.
• IPM demonstrated morphological differences: H cells had low nuclear OPD and ROS vesicles, while L cells had low cytoplasmic OPD and no ROS vesicles.
• L cells exhibited higher average optical path delay (OPD) and dry mass compared to H cells.
• IPM analysis of flowing neutrophils showed approximately 20% H cells in BBDs versus only 4% in MDS patients.

Conclusions:

• Interferometric phase microscopy (IPM) can differentiate neutrophil populations based on subtle morphological and biophysical properties.
• The reduced presence of H cells in MDS patients suggests IPM's potential as a diagnostic tool for myelodysplastic syndromes.
• This non-staining imaging technique offers a promising avenue for the rapid diagnosis of complex hematological disorders.