4.8 Article

Label-free hematology analysis using deep-ultraviolet microscopy

Publisher

NATL ACAD SCIENCES
DOI: 10.1073/pnas.2001404117

Keywords

deep-UV microscopy; molecular imaging; hematology analysis; point-of-care diagnosis; label-free cell classification

Funding

  1. Massner Lane Family Foundation
  2. Burroughs Wellcome Fund [CASI BWF 1014540]
  3. National Science Foundation [NSF CBET CAREER 1752011]
  4. Donaldson Charitable Trust Research Synergy Fund Award
  5. Winship Cancer Institute of Emory University
  6. Aflac Cancer & Blood Disorders Center at Children's Healthcare of Atlanta
  7. Wallace H. Coulter Biomedical Engineering Department at Emory University
  8. Wallace H. Coulter Biomedical Engineering Department at Georgia Institute of Technology

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Hematological analysis, via a complete blood count (CBC) and microscopy, is critical for screening, diagnosing, and monitoring blood conditions and diseases but requires complex equipment, multiple chemical reagents, laborious system calibration and procedures, and highly trained personnel for operation. Here we introduce a hematological assay based on label-free molecular imaging with deep-ultraviolet microscopy that can provide fast quantitative information of key hematological parameters to facilitate and improve hematological analysis. We demonstrate that this label-free approach yields 1) a quantitative five-part white blood cell differential, 2) quantitative red blood cell and hemoglobin characterization, 3) clear identification of platelets, and 4) detailed subcellular morphology. Analysis of tens of thousands of live cells is achieved in minutes without any sample preparation. Finally, we introduce a pseudocolorization scheme that accurately recapitulates the appearance of cells under conventional staining protocols for microscopic analysis of blood smears and bone marrow aspirates. Diagnostic efficacy is evaluated by a panel of hematologists performing a blind analysis of blood smears from healthy donors and thrombocytopenic and sickle cell disease patients. This work has significant implications toward simplifying and improving CBC and blood smear analysis, which is currently performed manually via bright-field microscopy, and toward the development of a low-cost, easy-to-use, and fast hematological analyzer as a point-of-care device and for low-resource settings.

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