Dataset Information

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

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.

SUBMITTER: Kaza N

PROVIDER: S-EPMC8353376 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Abnormally shaped red blood cells (RBCs), called poikilocytes, can cause anemia. At present, the biochemical abnormalities in poikilocytes are not well understood. Normal RBCs and poikilocytes were analyzed using whole-blood and single-cell methods. Poikilocytes were induced in rat blood by intragastrically administering titanium dioxide (TiO2) nanoparticles. Complete blood count and inductively coupled plasma mass spectrometry analyses were performed on whole-blood to measure average RBC morphology, blood hemoglobin (HGB), iron content, and other blood parameters. Follow-up confocal Raman spectroscopy was performed on single RBCs to analyze cell-type-specific HGB content. Two types of poikilocytes, acanthocytes and echinocytes, were observed in TiO2 blood samples, along with normal RBCs. Acanthocytes (diameter 7.7??±??0.5???m) and echinocytes (7.6??±??0.6???m) were microscopically larger (p??<??0.05) than normal RBCs (6.6??±??0.4???m) found in control blood samples (no TiO2 administration). Similarly, mean corpuscular volume was higher (p??<??0.05) in TiO2 whole-blood (70.70??±??1.97??fl) than in control whole-blood (67.42??±??2.03??fl). Poikilocytes also had higher HGB content. Mean corpuscular hemoglobin was higher (p??<??0.05) in TiO2 whole-blood (21.84??±??0.75??pg) than in control whole-blood (20.8??±??0.32??pg). Iron content was higher (p??<??0.001) in TiO2 whole-blood (697.0??±??24.5??mg??/??l) than in control whole-blood (503.4??±??38.5??mg??/??l), which supports elevated HGB as iron is found in HGB. HGB-associated Raman bands at 1637, 1585, and 1372??cm??-??1 had higher (p??<??0.001) amplitudes in acanthocytes and echinocytes than in RBCs from control blood and normal RBCs from TiO2 blood. Further, the 1585-cm??-??1 band had a lower (p??< ?0.05) amplitude in normal RBCs from TiO2 versus control RBCs. This represents biochemical abnormalities in normal appearing RBCs. Overall, poikilocytes, especially acanthocytes, have elevated HGB.