scholarly journals Accurate and independent measurement of volume and hemoglobin concentration of individual red cells by laser light scattering

Blood ◽  
1986 ◽  
Vol 68 (2) ◽  
pp. 506-513 ◽  
Author(s):  
N Mohandas ◽  
YR Kim ◽  
DH Tycko ◽  
J Orlik ◽  
J Wyatt ◽  
...  

Cell volume (MCV) and hemoglobin concentration (MCHC) are the red cell indices used to characterize the blood of patients with anemia. Since the introduction of flow cytometric methods for the measurement of these indices, it has generally been assumed that the values derived by these instruments are accurate. However, it has recently been shown that a number of cellular factors, including alterations in cellular deformability, can lead to inaccurate measurement of cell volume by these automated instruments. Because cell hemoglobin concentration and hematocrit are computed from the measured values of cell volume, accuracy of these indices is also compromised by inaccurate determination of cell volume. A recently developed experimental flow cytometric method based on laser light scattering, which can independently measure volume and hemoglobin concentration, has been used in the present study to measure MCV and MCHC of density- fractionated normal and sickle red cells, hydrated and dehydrated normal red cells, and various pathologic cells. We found that the new method accurately measures both volume and hemoglobin concentrations over a wide range of MCV (30 to 120 fL) and MCHC (27 to 45 g/dL) values. This is in contrast to currently available methods in which hemoglobin concentration values are accurately measured over a more limited range (27 to 35 g/dL). In addition, as the experimental method independently measures volume and hemoglobin concentration of individual red cells, it allowed us to generate histograms of volume and hemoglobin concentration distribution and derive coefficient of variation for volume distribution and standard deviation of hemoglobin concentration distribution. We have been able to document that volume and hemoglobin concentration distributions can vary independently of each other in pathologic red cell samples.

Blood ◽  
1986 ◽  
Vol 68 (2) ◽  
pp. 506-513 ◽  
Author(s):  
N Mohandas ◽  
YR Kim ◽  
DH Tycko ◽  
J Orlik ◽  
J Wyatt ◽  
...  

Abstract Cell volume (MCV) and hemoglobin concentration (MCHC) are the red cell indices used to characterize the blood of patients with anemia. Since the introduction of flow cytometric methods for the measurement of these indices, it has generally been assumed that the values derived by these instruments are accurate. However, it has recently been shown that a number of cellular factors, including alterations in cellular deformability, can lead to inaccurate measurement of cell volume by these automated instruments. Because cell hemoglobin concentration and hematocrit are computed from the measured values of cell volume, accuracy of these indices is also compromised by inaccurate determination of cell volume. A recently developed experimental flow cytometric method based on laser light scattering, which can independently measure volume and hemoglobin concentration, has been used in the present study to measure MCV and MCHC of density- fractionated normal and sickle red cells, hydrated and dehydrated normal red cells, and various pathologic cells. We found that the new method accurately measures both volume and hemoglobin concentrations over a wide range of MCV (30 to 120 fL) and MCHC (27 to 45 g/dL) values. This is in contrast to currently available methods in which hemoglobin concentration values are accurately measured over a more limited range (27 to 35 g/dL). In addition, as the experimental method independently measures volume and hemoglobin concentration of individual red cells, it allowed us to generate histograms of volume and hemoglobin concentration distribution and derive coefficient of variation for volume distribution and standard deviation of hemoglobin concentration distribution. We have been able to document that volume and hemoglobin concentration distributions can vary independently of each other in pathologic red cell samples.


Blood ◽  
1952 ◽  
Vol 7 (10) ◽  
pp. 1017-1019 ◽  
Author(s):  
D. C. VAN DYKE ◽  
J. F. GARCIA ◽  
M. E. SIMPSON ◽  
R. L. HUFF ◽  
A. N. CONTOPOULOS ◽  
...  

Abstract The anemia which follows hypophysectomy is apparently due to absence of the anterior lobe of the hypophysis as removal of the intermediate and posterior lobes did not change the hemoglobin concentration, hematocrit or volume of circulating red cells.


Improvement in bioprocess control will require development of monitors for a wide range of cell-growth and downstream-process parameters. These requirements are examined in terms of the development and application of biosensor devices and physical measurement techniques. Acoustic, dielectric and laser light scattering techniques are discussed primarily as monitors and analysers for biomass parameters. Developments with biosensor devices are discussed in terms of their application in membrane sampling-flow analysis probes and the potential of more direct biosensing principles.


1993 ◽  
Vol 265 (2) ◽  
pp. C562-C570 ◽  
Author(s):  
M. McManus ◽  
J. Fischbarg ◽  
A. Sun ◽  
S. Hebert ◽  
K. Strange

A simple and relatively inexpensive device utilizing laser light scattering for the study of volume regulatory behavior and membrane transport phenomena in cells cultured on or affixed to a rigid substrate is described in detail. Validation of the method is provided by study of cell types with known volume regulatory responses. The method we describe has numerous advantages over currently available techniques used to monitor cell volume changes. These advantages include 1) the ability to rapidly detect and quantify small cell volume changes on-line, 2) the ability to maintain natural cell morphology, cell surface contacts, and cell-to-cell interactions, 3) the ability to easily control solution temperature and gas and solute composition, and 4) the ability to perform multiple perturbations in a single experiment. The light-scattering system we describe can be modified to allow for simultaneous measurement of light-scattering signals and fluorescence emission from intracellular ion-sensitive probes and membrane potential dyes. In addition, our method may be useful for the study of apical and basolateral membrane transport processes in epithelial monolayer cell cultures.


1993 ◽  
Vol 324 ◽  
Author(s):  
C. Pickering ◽  
D.A.O. Hope ◽  
W.Y. Leong ◽  
D.J. Robbins ◽  
R. Greef

AbstractIn-situ dual-wavelength ellipsometry and laser light scattering have been used to monitor growth of Si/Si1−x,Gex heterojunction bipolar transistor and multi-quantum well (MQW) structures. The growth rate of B-doped Si0 8Ge0.2 has been shown to be linear, but that of As-doped Si is non-linear, decreasing with time. Refractive index data have been obtained at the growth temperature for x = 0.15, 0.20, 0.25. Interface regions ∼ 6-20Å thickness have been detected at hetero-interfaces and during interrupted alloy growth. Period-to-period repeatability of MQW structures has been shown to be ±lML.


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