Quantification of the Mean and Distribution of Hemoglobin Content in Normal Human Blood Using Cell Tracking Velocimetry

Kim, James; Gómez-Pastora, Jenifer; Gilbert, Christopher J.; Weigand, Mitchell; Walters, Nicole; Reátegui, Eduardo; Palmer, Andre F.; Yazer, Mark H.; Zborowski, Maciej; Chalmers, Jeffrey J.

doi:10.1021/acs.analchem.9b04302

Cited by 18 publications

(25 citation statements)

References 16 publications

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“…These graphs include the settling velocity, u s , versus the magnetic velocity, u m , scatter plot with appropriate histograms aligned along the side and top, respectively. The range in distribution presented here is consistent with what we have reported earlier in a study of 17 healthy donors (Kim et al 2020). Ongoing studies are investigating the significance of these distributions in healthy, anemic and patients with sickle cell disease.…”

Section: Reactions With Rbcs and Ctvsupporting

confidence: 91%

Magnetophoretic and Spectral Characterization of Oxyhemoglobin to Deoxyhemoglobin: Chemical vs Enzymatic Process

Weigand

Gómez-Pastora

Kim

et al. 2020

Preprint

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A new method for hemoglobin (Hb) deoxygenation and re-oxygenation, in suspension or within red blood cells, RBCs, is described using the commercial enzyme product, EC-Oxyrase®. This method using EC-Oxyrase has several advantages over established deoxygenation methodologies, such as avoiding side reactions that produce methemoglobin, eliminating the need of a sparging deoxygenation gas and airtight vessels, as well as easy re-oxygenation by washing and adding to a normal buffer with dissolved oxygen (DO). Spectra of deoxyHb and metHb from RBCs using three preparation methods: sodium dithionite, sodium nitrite and Oxyrase, show high purity of the deoxyHb using Oxyrase (with little to no methemoglobin or hemichrome production from side reactions). The deoxygenation action of Oxyrase follows first order reaction kinetics. Paramagnetic characteristics of intracellular hemoglobin in RBCs are compared using cell tracking velocimetry for healthy and sickle cell disease (SCD) donors and oxygen dissociation curves show that the function of healthy RBCs is unchanged after Oxyrase treatment. The results confirm that this enzymatic approach to deoxygenation produces pure deoxyhemoglobin, can be re-oxygenated easily, prepared aerobically and has similar paramagnetic mobility to the existing methods.

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Section: Reactions With Rbcs and Ctvsupporting

confidence: 91%

Magnetophoretic and Spectral Characterization of Oxyhemoglobin to Deoxyhemoglobin: Chemical vs Enzymatic Process

Weigand

Gómez-Pastora

Kim

et al. 2020

Preprint

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“…By observing both Figure 4 and Table 1, it can be concluded that the MCH (expressed in pg of Hb per RBC) values reported by CTV are within the normal range reported for healthy individuals, between 27 and 33 pg. Moreover, our previous study assesed the performance of CTV on measuring mass of Hb on indicidual RBCs by comparing CTV results to the standard method based on spectrophotometry, and our instrument reported accurate measurements/analysis [21]. Nevertheless, when comparing the MCHC measured by CTV to the reference range of 28-41 g/dL [23], CTV measured higher MCHC due to the fact that the measured MCV is smaller than the reference range, as happened to the RDW values from the CTV instrument.…”

Section: Hemoglobin Mass and Concentration In Individual Rbcsmentioning

confidence: 86%

“…Total hemoglobin, Hb, is measured from whole blood samples by using the spectrophotometric method commonly employed in clinical laboratories, as described in our previous work [21]. Briefly, the samples obtained for spectrophotometric Hb concentration determination were diluted (4X) in deionized water in order to lyse the RBCs.…”

Section: Spectrophotometric Analysismentioning

confidence: 99%

“…The calibration and operating procedure for using CTV to magnetically characterize RBCs has been described in previous reports [21,22]. Supplementary Figures S1 and S2 report the working principle and the optimization of the CTV operation procedure.…”

Section: Ctv Analysismentioning

confidence: 99%

“…Previously, the relationship between the magnetic and settling velocity and the amount of hemoglobin in the RBC, accounting for volume, has been described in detail [20,21]:…”

Section: Introductionmentioning

confidence: 99%

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Potential of Cell Tracking Velocimetry as an Economical and Portable Hematology Analyzer

Gómez-Pastora

Kim

Weigand

et al. 2021

Preprint

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Anemia and iron deficiency continue to be the most prevalent nutritional disorders in the world, affecting billions of people in both developed and developing countries. The initial diagnosis of anemia is typically based on several markers, including red blood cell (RBC) count, hematocrit and total hemoglobin. Using modern hematology analyzers, erythrocyte parameters such as mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), etc. are also being used. However, most of these commercially available analyzers pose several disadvantages: they are expensive instruments that require significant bench space and are heavy enough to limit their use to a specific lab and leading to a delay in results, making them less practical as a point-of-care instrument that can be used for swift clinical evaluation. Thus, there is a need for a portable and economical hematology analyzer that can be used at the point of need. In this work, we evaluated the performance of a system referred to as the cell tracking velocimetry (CTV) to measure several hematological parameters from fresh human blood obtained from healthy donors. Our system, based on the paramagnetic behavior that methemoglobin containing RBCs experience when suspended in water after applying a magnetic field, uses a combination of magnets and microfluidics and has the ability to track the movement of thousands of red cells in a short period of time. This allows us to measure not only traditional RBC indices but also novel parameters that are only available for analyzers that assess erythrocytes on a cell by cell basis. As such, we report, for the first time, the use of our CTV as a hematology analyzer that is able to measure red cell volume or MCV, red cell hemoglobin mass or MCH, hemoglobin concentration (MCHC), red cell distribution width (RDW) and the percentage of hypochromic cells, which is an indicator of insufficient marrow iron supply that reflects recent iron reduction. Our initial results indicate that most of the parameters measured with CTV are within the normal range for healthy adults. Only the parameters related to the red cell volume (primarily MCV and RDW) were outside the normal range. We observed significant discrepancies between the MCV measured by our technology (and also by an automated cell counter) and the manual MCV measured through the hematocrit obtained by packed cell volume method, which are attributed to the artifacts of plasma trapping and cell shrinkage. While there may be limitations for measuring MCV, this device offers a novel point of care instrument to provide rapid RBC parameters such as iron stores that are otherwise not rapidly available to the clinician. Thus, our CTV is a promising technology with the potential to be employed as an accurate, economical, portable and fast hematology analyzer after applying instrument-specific reference ranges or correction factors.

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Iron in blood cells: Function, relation to disease, and potential for magnetic separation

Barua

Ciannella

Tijani

et al. 2023

Biotech & Bioengineering

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Iron in blood cells has several physiological functions like transporting oxygen to cells and maintaining iron homeostasis. Iron is primarily contained in red blood cells (RBCs), but monocytes also store iron as these cells are responsible for the recycling of senescent RBCs. Iron also serves an important role related to the function of different leukocytes. In inflammation, iron homeostasis is dependent on cytokines derived from T cells and macrophages. Fluctuations of iron content in the body lead to different diseases. Iron deficiency, which is also known as anemia, hampers different physiological processes in the human body. On the other hand, genetic or acquired hemochromatosis ultimately results in iron overload and leads to the failure of different vital organs. Different diagnoses and treatments are developed for these kinds of disorders, but the majority are costly and suffer from side effects. To address this issue, magnetophoresis could be an attractive technology for the diagnosis (and in some cases treatment) of these pathologies due to the paramagnetic character of the cells containing iron. In this review, we discuss the main functions of iron in blood cells and iron‐related diseases in humans and highlight the potential of magnetophoresis for diagnosing and treating some of these disorders.

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Quantification of the Mean and Distribution of Hemoglobin Content in Normal Human Blood Using Cell Tracking Velocimetry

Cited by 18 publications

References 16 publications

Magnetophoretic and Spectral Characterization of Oxyhemoglobin to Deoxyhemoglobin: Chemical vs Enzymatic Process

Magnetophoretic and Spectral Characterization of Oxyhemoglobin to Deoxyhemoglobin: Chemical vs Enzymatic Process

Potential of Cell Tracking Velocimetry as an Economical and Portable Hematology Analyzer

Iron in blood cells: Function, relation to disease, and potential for magnetic separation

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