Label-Free High-Throughput Leukemia Detection by Holographic Microscopy

Area of Science:

• Biomedical Engineering
• Flow Cytometry
• Digital Holographic Microscopy

Background:

• Complete blood count and leukocyte differentiation (DIFF) are essential diagnostic tests.
• Current DIFF methods often require reagents and can be complex.

Purpose of the Study:

• To develop a label-free, flow cytometry-based method for leukocyte differentiation.
• To utilize digital holographic microscopy for enhanced morphological analysis of leukocytes.

Main Methods:

• Employed digital holographic microscopy with 2D hydrodynamic focusing for label-free leukocyte imaging.
• Applied Principal Component Analysis (PCA) to morphological characteristics of reconstructed images for classification.
• Validated the method on 20 blinded clinical samples.

Main Results:

• Successfully classified nine leukocyte types and identified different leukemia types.
• Demonstrated the ability to detect the disappearance of acute myeloid leukemia cells during remission.
• Achieved 70% correct classification and 20% near-correct classifications in blinded samples.

Conclusions:

• The developed flow cytometry method offers automated and reagent-free diagnosis of hematological disorders.
• Digital holographic microscopy provides rich phase contrast for accurate leukocyte morphology analysis.
• The technique shows broad clinical applicability for hematological diagnostics.