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Direct Quantification of Single Red Blood Cell Hemoglobin Concentration with Multiphoton Microscopy
Andrew T Francis1, Melanie J Shears2, Sean C Murphy2
1Department of Chemistry, University of Washington, Seattle, Washington 98195, United States.
Analytical Chemistry
|August 14, 2020
Summary
This study introduces a new imaging method to measure hemoglobin in single red blood cells (RBCs). This technique quantifies mean corpuscular hemoglobin concentration (MCHC) for improved disease diagnosis and treatment.
Area of Science:
- Biomedical Optics
- Hematology
- Parasitology
Background:
- Blood disorders significantly alter red blood cell (RBC) characteristics.
- Mean corpuscular hemoglobin concentration (MCHC) is a key diagnostic marker.
- Current MCHC analysis uses bulk measurements, missing cellular heterogeneity.
Purpose of the Study:
- To develop a novel multimodal imaging technique for single-RBC MCHC quantification.
- To enable precise measurement of hemoglobin concentration at the single-cell level.
- To investigate RBC heterogeneity in health and disease states.
Main Methods:
- Utilized a multimodal multiphoton approach combining stimulated Raman scattering (SRS) for water and transient absorption (TA) for hemoglobin.
- Implemented a dual-channel lock-in detection with orthogonal modulation for simultaneous imaging.
- Employed water as an internal standard for accurate MCHC quantification.
Main Results:
- Successfully quantified MCHC at single RBC resolution.
- Demonstrated the capability to differentiate MCHC in healthy RBCs versus those infected with *Plasmodium yoelii*.
- Validated the multimodal imaging approach for pathological analysis.
Conclusions:
- The novel multimodal imaging technique provides high-resolution MCHC measurements.
- This method offers a powerful tool for studying RBC heterogeneity and parasitic infections.
- Potential for improved diagnosis and therapeutic monitoring of blood-related diseases.
