Hemoglobin quantification in red blood cells via dry mass mapping based on UV absorption

• Type of Material: Article
• Authors: Nischita Kaza, Georgia Institute of Technology; Ashkan Ojaghi, Georgia Institute of Technology; Francisco Robles, Emory University
• Language: English
• Date: 2021-08-01
• Publisher: SPIE-SOC PHOTO-OPTICAL INSTRUMENTATION ENGINEERS
• Publication Version: Final Published Version
• Copyright Statement: © 2021 The Authors
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1117/1.JBO.26.8.086501
• Title of Journal or Parent Work: JOURNAL OF BIOMEDICAL OPTICS
• Volume: 26
• Issue: 8
• Keywords: red blood cells; mass mapping; quantitative phase imaging; INTRACELLULAR PROTEIN; MICROSCOPY; Biochemical Research Methods; hemoglobin; CYTOMETRY; Optics; Science & Technology; NUCLEIC-ACID; VOLUME; deep-UV microscopy; hematology analysis; Radiology, Nuclear Medicine & Medical Imaging; Physical Sciences; Biochemistry & Molecular Biology; QUANTITATIVE PHASE; Life Sciences & Biomedicine; DYNAMICS; NEUTROPHIL LEFT SHIFT
• Research Categories: Engineering, Electronics and Electrical; Engineering, Biomedical

Abstract

SIGNIFICANCE: The morphological properties and hemoglobin (Hb) content of red blood cells (RBCs) are essential biomarkers to diagnose or monitor various types of hematological disorders. Label-free mass mapping approaches enable accurate Hb quantification from individual cells, serving as promising alternatives to conventional hematology analyzers. Deep ultraviolet (UV) microscopy is one such technique that allows high-resolution, molecular imaging, and absorption-based mass mapping. AIM: To compare UV absorption-based mass mapping at four UV wavelengths and understand variations across wavelengths and any assumptions necessary for accurate Hb quantification. APPROACH: Whole blood smears are imaged with a multispectral UV microscopy system, and the RBCs' dry masses are computed. This approach is compared to quantitative phase imaging-based mass mapping using data from an interferometric UV imaging system. RESULTS: Consistent Hb mass and mean corpuscular Hb values are obtained at all wavelengths, with the precision of the single-cell mass measurements being nearly identical at 220, 260, and 280 nm but slightly lower at 300 nm. CONCLUSIONS: A full hematological analysis (including white blood cell identification and characterization, and Hb quantification) may be achieved using a single UV illumination wavelength, thereby improving the speed and cost-effectiveness.