The Effects of RPM and Recycle on Separation Efficiency in a Clinical Blood Cell Centrifuge

A COBE blood cell centrifuge, model 2997 with a single stage channel, was modified to allow computer controlled sampling, and to allow recycle of red blood cells (RBCs) and plasma streams using bovine whole blood. The effects of recycle of the packed RBC and plasma product streams, and of the centrifuge RPM on platelet and white blood cell (WBC) separation efficiencies were quantified using a central composite factorial experimental design. These data were then fit using second order models. Both the model for the WBC separation efficiency and the model for the platelet separation efficiency predict that RPM has the greatest effect on separation efficiency and that RBC and plasma recycle have detrimental effects at moderate to low RPM, but have negligible impact on separation efficiency at high RPM.

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IN VITRO INTERACTIONS BETWEEN OXYGEN AND CARBON DIOXIDE TRANSPORT IN THE BLOOD OF THE SEA LAMPREY (PETROMYZON MARINUS)

Journal of Experimental Biology ◽

10.1242/jeb.173.1.25 ◽

1992 ◽

Vol 173 (1) ◽

pp. 25-41 ◽

Cited By ~ 1

Author(s):

R. A. Ferguson ◽

N. Sehdev ◽

B. Bagatto ◽

B. L. Tufts

Keyword(s):

Carbon Dioxide ◽

Blood Cell ◽

Red Blood Cells ◽

Red Blood Cell ◽

Whole Blood ◽

Blood Cells ◽

Sea Lamprey ◽

Carbon Dioxide Transport ◽

Dissociation Curves

In vitro experiments were carried out to examine the interactions between oxygen and carbon dioxide transport in the blood of the sea lamprey. Oxygen dissociation curves for whole blood obtained from quiescent lampreys had Hill numbers (nH) ranging from 1.52 to 1.89. The Bohr coefficient for whole blood was -0.17 when extracellular pH (pHe) was considered, but was much greater (-0.63) when red blood cell pH (pHi) was considered. The pHi was largely dependent on haemoglobin oxygen- saturation (SO2) and the pH gradient across the red blood cell membrane was often reversed when PCO2 was increased and/or SO2 was lowered. The magnitude of the increase in pHi associated with the Haldane effect ranged from 0.169 pH units at 2.9 kPa PCO2 to 0.453 pH units at a PCO2 of 0.2 kPa. Deoxygenated red blood cells had a much greater total CO2 concentration (CCO2) than oxygenated red blood cells, but the nonbicarbonate buffer value for the red blood cells was unaffected by oxygenation. Plasma CCO2 was not significantly different under oxygenated or deoxygenated conditions. Partitioning of CO2 carriage in oxygenated and deoxygenated blood supports recent in vivo observations that red blood cell CO2 carriage can account for much of the CCO2 difference between arterial and venous blood. Together, the results also suggest that oxygen and carbon dioxide transport may not be tightly coupled in the blood of these primitive vertebrates. Finally, red cell sodium concentrations were dependent on oxygen and carbon dioxide tensions in the blood, suggesting that sodium-dependent ion transport processes may contribute to the unique strategy for gas transport in sea lamprey blood.

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RED BLOOD CELL LYSIS MAY INFLUENCE PLATELET AGGREGATION IN WHOLE BLOOD AGGREGOMETER

10.1055/s-0038-1644552 ◽

1987 ◽

Author(s):

A G Herman ◽

H Bult

Keyword(s):

Platelet Aggregation ◽

Blood Cell ◽

Red Blood Cells ◽

Red Blood Cell ◽

Whole Blood ◽

Blood Cells ◽

Adenine Nucleotide ◽

Cell Lysis ◽

Blood Collection ◽

Adenosine Uptake

The electronic whole blood aggregometer (WBA) has the advantage that it enables the study of platelet aggregation in whole blood shortly after blood collection. Using the WBA varying results have been obtained with respect to the anti-aggregating activity of dipyridamole. As dipyridamole is an efficient inhibitor of adenosine uptake, we tested whether the degree of red blood cell lysis (and thus availability of adenine nucleotides) affected its efficacy. Citrate (10.7 mM) blood was stored in sealed tubes and used between 20 and 100 min after venipuncture. One ml was placed in a Chronolog Model 540 WBA together with 10 μl 0.9 % NaCl, dipyridamole (final conc. 3, 10 or 30 μM) or its solvent (final conc. 0.03, 0.1 or 0.3 %). After reaching a stable baseline and WBA calibration, aggregation was induced by injection of 10 μM ADP dissolved in 10 μl 0.9 % NaCl (one channel) or 10 μl distilled water (other channel). Maximum impedance increase in 10 min was measured, red blood cells were removed by centrifugation, and from microhematocrit and absorbance at 416 nm the volume of lysed packed red blood cells was estimated. ADP caused aggregation (12.9 ± 1.9 and 11.1 ± 0.8, Ohm) and there was red blood cell lysis (2.8 ± 0.5 and 0.8 ± 0.2 μl red blood cells, ADP resp. in H20 and 0.9 % NaCF, n = 6). Dipyridamole (30 μM) suppressed aggregation when compared with solvent, but only when ADP was given in H20 (reduction resp. 4.4 ± 1.4 and 1.9 ± 1.6 ohm). Moreover, there was a negative correlation between the degree of haemolysis and the aggregation response at 10 as well as 30 μM dipyridamole. This reduced aggregation with increasing haemolysis was not observed in the presence of the corresponding solvent concentrations. The red blood cell lysis was proportional to the plasma ATP (luciferine-luciferase method) concentration as an index of adenine nucleotide leakage. In conclusion, a certain degree of haemolysis caused by stirring and injection with a microsyringe is inherent to the WBA, but use of hypotonic vehicles should be avoided. Release of red blood cell constituents may affect platelet aggregation as such, or interfere with the activity of adenosine uptake inhibitors and possibly other drugs. It may help to explain some of the variable results obtained with dipyridamole.

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An Evaluation of Morphological Changes and Deformability of Suspended Red Blood Cells Prepared Using Whole Blood with Different Hemoglobin Levels of Tibetans

Transfusion Medicine and Hemotherapy ◽

10.1159/000513319 ◽

2021 ◽

pp. 1-10

Author(s):

Rui Zhong ◽

Dingding Han ◽

Xiaodong Wu ◽

Hong Wang ◽

Wanjing Li ◽

...

Keyword(s):

Red Blood Cells ◽

Whole Blood ◽

Blood Cells ◽

Morphological Changes ◽

Osmotic Fragility ◽

Spherical Shape ◽

Hypoxic Environment ◽

Wide Range ◽

Group A ◽

Cell Membrane Protein

Background: The hypoxic environment stimulates the human body to increase the levels of hemoglobin (HGB) and hematocrit and the number of red blood cells. Such enhancements have individual differences, leading to a wide range of HGB in Tibetans’ whole blood (WB). Study Design: WB of male Tibetans was divided into 3 groups according to different HGB (i.e., A: >120 but ≤185 g/L, B: >185 but ≤210 g/L, and C: >210 g/L). Suspended red blood cells (SRBC) processed by collected WB and stored in standard conditions were examined aseptically on days 1, 14, 21, and 35 after storage. The routine biochemical indexes, deformability, cell morphology, and membrane proteins were tested. Results: Mean corpuscular volume, adenosine triphosphate, pH, and deformability were not different in group A vs. those in storage (p > 0.05). The increased rate of irreversible morphology of red blood cells was different among the 3 groups, but there was no difference in the percentage of red blood cells with an irreversible morphology after 35 days of storage. Group C performed better in terms of osmotic fragility and showed a lower rigid index than group A. Furthermore, SDS-PAGE revealed similar cross-linking degrees of cell membrane protein but the band 3 protein of group C seemed to experience weaker clustering than that of group A as detected by Western Blot analysis after 35 days of storage. Conclusions: There was no difference in deformability or morphological changes in the 3 groups over the 35 days of storage. High HGB levels of plateau SRBC did not accelerate the RBC change from a biconcave disc into a spherical shape and it did not cause a reduction in deformability during 35 days of preservation in bank conditions.

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Glycophorin A-based exclusion of red blood cells for flow cytometric analysis of platelet glycoprotein expression in citrated whole blood

Clinical Chemistry and Laboratory Medicine (CCLM) ◽

10.1515/cclm-2020-0014 ◽

2020 ◽

Vol 0 (0) ◽

Author(s):

Christina Berens ◽

Johannes Oldenburg ◽

Bernd Pötzsch ◽

Jens Müller

Keyword(s):

Red Blood Cells ◽

Expression Analysis ◽

Whole Blood ◽

Blood Cells ◽

Control Sample ◽

Small Sample ◽

Platelet Glycoprotein ◽

Glycophorin A ◽

Giant Platelets ◽

Platelet Counts

AbstractObjectivesAnalysis of platelet glycoprotein (GP) expression by flow cytometry is applied for diagnostic confirmation of GP-associated thrombocytopathies. While platelet-rich plasma may be used for distinct identification of target events, this strategy is not feasible for small sample volumes or for patients showing low platelet counts and/or giant platelets. However, also the use of whole blood (WB) is hampered by the difficulty to discriminate platelets from red blood cells (RBC) in such patients. To circumvent these limitations, we evaluated the feasibility of a RBC gating-out strategy.MethodsIn addition to platelet GPIb, GPIIa/IIIa, as well as P-selectin (CD62P), citrated whole blood (CWB) samples were stained for RBC-specific glycophorin A (CD235a). CD235a-negative platelet events were further discriminated by forward-/side-scatter characteristics and platelet GP expressions analyzed relative to that of a healthy control sample processed in parallel.ResultsEstablished reference intervals allowed for clear identification of decreased GPIIb/IIIa- or GPIb expression pattern in samples of patients with confirmed Glanzmann thrombasthenia or Bernard–Soulier syndrome, respectively. It could be shown that the analysis of 2,500 platelet events is sufficient for reliable GP expression analysis, rendering the proposed method applicable to samples with low platelet counts.ConclusionsThis study demonstrates the feasibility of CD235a-based exclusion of RBC for platelet GP expression analysis in CWB. In contrast to direct staining of platelet-specific antigens for target identification, this indirect gating out approach is generally applicable independent of any underlying platelet GP expression deficiency.

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Microfluidic electrical impedance assessment of red blood cell mediated microvascular occlusion

Lab on a Chip ◽

10.1039/d0lc01133a ◽

2021 ◽

Author(s):

Yuncheng Man ◽

Debnath Maji ◽

Ran An ◽

Sanjay Ahuja ◽

Jane A Little ◽

...

Keyword(s):

Sickle Cell Disease ◽

Blood Cell ◽

Red Blood Cells ◽

Sickle Cell ◽

Red Blood Cell ◽

Blood Cells ◽

Electrical Impedance ◽

Cell Disease ◽

Blood Disorders

Alterations in the deformability of red blood cells (RBCs), occurring in hemolytic blood disorders such as sickle cell disease (SCD), contributes to vaso-occlusion and disease pathophysiology. However, there are few...

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