Flow-cytometric light scattering measurement of red blood cell volume and hemoglobin concentration

Abstract Circulating red blood cell volumes were determined by the carbon monoxide method, and plasma volumes were calculated in four men 20, 29, 71, and 75 years old, and two women 29 years of age before, during, and after exposure to an altitude of 3800 m. In the four youngest subjects there were early increases in hemoglobin concentration during the first days at the stated altitude attributed to decreases in plasma volume. At the same time, hemoglobin concentration decreased and plasma volume increased in the oldest subject. Red cell volumes were slow to change, and it was concluded that 3 weeks or more of exposure to this altitude are required to affect significantly the red cell volume in man.

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Why is there no carbonic anhydrase activity available to fish plasma?

Journal of Experimental Biology ◽

10.1242/jeb.198.1.31 ◽

1995 ◽

Vol 198 (1) ◽

pp. 31-38 ◽

Cited By ~ 2

Author(s):

J Lessard ◽

A Val ◽

S Aota ◽

D Randall

Keyword(s):

Carbonic Anhydrase ◽

Blood Cell ◽

Cell Volume ◽

Red Blood Cell ◽

Short Circuit ◽

Hemoglobin Concentration ◽

Carbonic Anhydrase Activity ◽

Control Group ◽

Acid Loading

Carbonic anhydrase (CA) is absent from the plasma of vertebrates. In vitro, CA in fish plasma will short-circuit the effect of catecholamines, which is to increase red blood cell (RBC) pH and volume, both of which enhance the affinity of hemoglobin for O2. CA was infused into trout for a period of 6 h and injected after 48 h, during which the animal was submitted to deep hypoxia (PO2=3035 mmHg; 4.04.7 kPa). O2 content, lactate content, catecholamine levels, hematocrit, hemoglobin concentration and pHi were similar to those in the saline-infused control group. In contrast, cell volume was significantly higher and pHe, total CO2 content and organic phosphate levels were significantly lower than in the control group. The concentration of CA was not high enough completely to short-circuit the increase in pHi and red blood cell volume caused by catecholamines. The lower pHe in the CA-infused animals could enhance the activity of the Na+/H+ pump, which would keep the nucleotide triphosphate levels low. pH is a balance between acid loading at the muscle and acid excretion at the gills or the kidneys; we cannot distinguish between which of these resulted in a decrease of plasma pH. In conclusion, CA in plasma did not cause the expected reduction in blood oxygen content but did have a marked effect on plasma total CO2 content.

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Alterations in Red Blood Cell Volume and Hemoglobin Concentration, Viscoelastic Properties, and Mechanical Fragility Caused by Continuous Flow Pumping in Calves

Artificial Organs ◽

10.1111/j.1525-1594.2011.01317.x ◽

2011 ◽

Vol 35 (8) ◽

pp. 791-799 ◽

Cited By ~ 4

Author(s):

Naoyuki Yokoyama ◽

Daisuke Sakota ◽

Eiki Nagaoka ◽

Setsuo Takatani

Keyword(s):

Blood Cell ◽

Cell Volume ◽

Red Blood Cell ◽

Continuous Flow ◽

Viscoelastic Properties ◽

Hemoglobin Concentration

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

Blood ◽

10.1182/blood.v68.2.506.bloodjournal682506 ◽

1986 ◽

Vol 68 (2) ◽

pp. 506-513 ◽

Cited By ~ 1

Author(s):

N Mohandas ◽

YR Kim ◽

DH Tycko ◽

J Orlik ◽

J Wyatt ◽

...

Keyword(s):

Light Scattering ◽

Cell Volume ◽

Concentration Distribution ◽

Laser Light ◽

Hemoglobin Concentration ◽

Laser Light Scattering ◽

Red Cells ◽

Red Cell ◽

Flow Cytometric ◽

Wide Range

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.

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Regulation of blood volume in lowlanders exposed to high altitude

Journal of Applied Physiology ◽

10.1152/japplphysiol.00118.2017 ◽

2017 ◽

Vol 123 (4) ◽

pp. 957-966 ◽

Cited By ~ 34

Author(s):

Christoph Siebenmann ◽

Paul Robach ◽

Carsten Lundby

Keyword(s):

Blood Cell ◽

High Altitude ◽

Cell Volume ◽

Blood Volume ◽

Plasma Volume ◽

Red Blood Cell ◽

Hemoglobin Concentration ◽

Circulating Blood Volume ◽

Oxyhemoglobin Saturation ◽

Physiological Relevance

Humans ascending to high altitude (HA) experience a reduction in arterial oxyhemoglobin saturation and, as a result, arterial O2content ([Formula: see text]). As HA exposure extends, this reduction in [Formula: see text] is counteracted by an increase in arterial hemoglobin concentration. Initially, hemoconcentration is exclusively related to a reduction in plasma volume (PV), whereas after several weeks a progressive expansion in total red blood cell volume (RCV) contributes, although often to a modest extent. Since the decrease in PV is more rapid and usually more pronounced than the expansion in RCV, at least during the first weeks of exposure, a reduction in circulating blood volume is common at HA. Although the regulation of hematological responses to HA has been investigated for decades, it remains incompletely understood. This is not only related to the large number of mechanisms that could be involved and the complexity of their interplay but also to the difficulty of conducting comprehensive experiments in the often secluded HA environment. In this review, we present our understanding of the kinetics, the mechanisms and the physiological relevance of the HA-induced reduction in PV and expansion in RCV.

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

Blood ◽

10.1182/blood.v68.2.506.506 ◽

1986 ◽

Vol 68 (2) ◽

pp. 506-513 ◽

Cited By ~ 123

Author(s):

N Mohandas ◽

YR Kim ◽

DH Tycko ◽

J Orlik ◽

J Wyatt ◽

...

Keyword(s):

Light Scattering ◽

Cell Volume ◽

Concentration Distribution ◽

Laser Light ◽

Hemoglobin Concentration ◽

Laser Light Scattering ◽

Red Cells ◽

Red Cell ◽

Flow Cytometric ◽

Wide Range

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.

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