cell tracking velocimetry
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PLoS ONE ◽  
2021 ◽  
Vol 16 (9) ◽  
pp. e0257061
Author(s):  
Mitchell R. H. Weigand ◽  
Jenifer Gómez-Pastora ◽  
James Kim ◽  
Matthew T. Kurek ◽  
Richard J. Hickey ◽  
...  

A new method for hemoglobin (Hb) deoxygenation, in suspension or within red blood cells (RBCs) is described using the commercial enzyme product, EC-Oxyrase®. The enzymatic deoxygenation method has several advantages over established deoxygenation methodologies, such as avoiding side reactions that produce methemoglobin (metHb), thus eliminating the need for an inert deoxygenation gas and airtight vessel, and facilitates easy re-oxygenation of Hb/RBCs by washing with a buffer that contains dissolved oxygen (DO). The UV-visible spectra of deoxyHb and metHb purified from human RBCs using three different preparation methods (sodium dithionite [to produce deoxyHb], sodium nitrite [to produce metHb], and EC-Oxyrase® [to produce deoxyHb]) show the high purity of deoxyHb prepared using EC-Oxyrase® (with little to no metHb or hemichrome production from side reactions). The oxyHb deoxygenation time course of EC-Oxyrase® follows first order reaction kinetics. The paramagnetic characteristics of intracellular Hb in RBCs were compared using Cell Tracking Velocimetry (CTV) for healthy and sickle cell disease (SCD) donors and oxygen equilibrium curves show that the function of healthy RBCs is unchanged after EC-Oxyrase® treatment. The results confirm that this enzymatic approach to deoxygenation produces pure deoxyHb, can be re-oxygenated easily, prepared aerobically and has similar paramagnetic mobility to existing methods of producing deoxyHb and metHb.


2021 ◽  
Author(s):  
Jenifer Gómez-Pastora ◽  
James Kim ◽  
Mitchell Weigand ◽  
Andre F. Palmer ◽  
Mark Yazer ◽  
...  

Abstract 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.


Author(s):  
Jenifer Gómez-Pastora ◽  
James Kim ◽  
Victor Multanen ◽  
Mitchell Weigand ◽  
Nicole Walters ◽  
...  

The presence of iron in circulating monocytes is well known as they play an essential role in iron recycling. It has been demonstrated that the iron content of blood cells can be measured through their magnetic behavior; however, the magnetic properties of different monocyte subtypes remain unknown. In this study, we report for the first time, the magnetic behavior of classical, intermediate and non-classical monocytes, which is related to their iron storage capacity. The magnetic properties of monocytes were compared to other blood cells, such as lymphocytes and red blood cells in the oxyhemoglobin and methemoglobin states, and a cancer cell type. For this analysis, we used an instrument referred to as Cell Tracking Velocimetry (CTV), which quantitatively characterizes the magnetic behavior of biological entities. Our results demonstrate that significant fractions of the intermediate and non-classical monocytes have high magnetophoretic mobilities, equivalent to methemoglobin red blood cells and higher than the classical subset, suggesting their higher iron storage capacities. Moreover, our findings have implications for the immunomagnetic separation industry; we demonstrate that negative magnetic isolation techniques for recovering monocytes from blood should be used with caution, as it is possible to lose magnetic monocytes when using this technique.


2019 ◽  
Vol 92 (2) ◽  
pp. 1956-1962 ◽  
Author(s):  
James Kim ◽  
Jenifer Gómez-Pastora ◽  
Christopher J. Gilbert ◽  
Mitchell Weigand ◽  
Nicole A. Walters ◽  
...  

The Analyst ◽  
2008 ◽  
Vol 133 (12) ◽  
pp. 1767 ◽  
Author(s):  
Xiaoxia Jin ◽  
Yang Zhao ◽  
Aaron Richardson ◽  
Lee Moore ◽  
P. Stephen Williams ◽  
...  

Author(s):  
Chang-Beom Kim ◽  
Edward Steager ◽  
Min Jun Kim

A tracking algorithm was developed to study the velocity field of flagellated bacteria, Serratia marcescens, swarming on a soft agar plate. Average velocities for local regions regularly arranged over the entire flow field were investigated rather than those for individual bacterium. The velocity field of the bacteria typically featured the combination of curvilinear translation and vortex modes. They repeated these patterns for a short time period, forming several groups and dissipating. To further investigate the flow patterns generated by the collective motion of the swarming bacteria, the velocity field on the swarm was spatially correlated.


2005 ◽  
Vol 66A (2) ◽  
pp. 103-108 ◽  
Author(s):  
Diane R. Leigh ◽  
Steffen Steinert ◽  
Lee R. Moore ◽  
Jeffrey J. Chalmers ◽  
Maciej Zborowski

The Analyst ◽  
2005 ◽  
Vol 130 (4) ◽  
pp. 514 ◽  
Author(s):  
Huading Zhang ◽  
Lee R. Moore ◽  
Maciej Zborowski ◽  
P. Stephen Williams ◽  
Shlomo Margel ◽  
...  

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