Cell-membrane and rheological mechanisms: dynamic osmotic hemolysis of human erythrocytes and repair of ghosts, as studied by resistive pulse spectroscopy

Yee, James P.; Mel, Howard C.

doi:10.3233/bir-1978-153-419

Cited by 25 publications

(11 citation statements)

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“…Figure 2 shows data from our laboratory on osmotic volume changes of erythrocytes subjected t o different concentrations of phosphate-buffered saline (PBS) solutions. These are plotted, for comparison, in the same normalized form as that used for Figure 1 fitted by the curve correspond to the raw, "measured" volumes of red cells, as directly obtained under "slowflow" conditions through the orifice (about 1 dsec), using resistive pulse spectroscopy (RPS), an extension of the Coulter counter methodology (Me1 and Yee, 1975;Yee and Mel, 1978). The solid points and associated straight line represent the "true" volumes, after having quantitatively corrected the "measured' volumes for shape factor, according to the method of Richieri et al (1985a,b).…”

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confidence: 99%

Alternative interpretation for the osmotic response of human erythrocytes

Massaldi

Richieri

Mel

1986

Journal Cellular Physiology

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In a recent publication, Heubusch et al. (J Cell. Physiol, 122:266-272, 1985) reported changes of erythrocyte volume measured by the Coulter counter technique over a wide range of osmolalities (160 to 3000 m0sm). Their results showed a partially hindered, nonlinear response, in contrast to classical observations made over more restricted osmolality ranges, using other methods. The authors suggested the underlying cause of this behavior to be a mechanical resistance of the membrane cytoskeleton. In this paper, we wish to offer a different interpretation of their results on erythrocyte osmotic behavior, based on similar experiments carried out in our laboratory, and supported by previous analyses from the literature. In particular, it is shown that the shape-factor correction to the electronic sizing measurement can correctly account for the observed deviations from linearity in the hypotonic range. In contrast, increased chemical nonideality and eventual hemolysis are the likely factors responsible for the behavior in the hypertonic range.

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confidence: 99%

Alternative interpretation for the osmotic response of human erythrocytes

Massaldi

Richieri

Mel

1986

Journal Cellular Physiology

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“…The volume increase is of the same magnitude when measured either from GA-fixed cells or from SEM micrographs. The maximal volume increase of 60-62",, in 0.34",, NaCl is smaller than the 78",, measured by a haematocrit method (Seeman et al, 1969), 74",, measured on an interference microscope (Evans & Fung, 1972) or 67",, measured on a resistive pulse spectroscope (Yee & Mel, 1978).…”

Section: Quantitative Resultsmentioning

confidence: 74%

“…The use of scanning electron microscopy (SEM) has yielded a better understanding of the shapes of red blood cells induced by ageing (Chailley et al, 1973), drugs (Sheetz & Singer, 1974;Fujii et al, 1979) or various osmotic (Metz et al, 1971;Bleau et al, 1975;Yee & Mel, 1978) or pathological conditions (Bessis & Weed, 1972;Kupari-Koby, 1976). Both SEM and high-voltage electron microscopy can also be used when visualizing the ultrastructure of haemolysed erythrocytes ('ghosts') (Sprandel et al, 1981;Wise et al, 1981), but SEM has seldom been used in quantitative studies on red blood cell geometry, such as the determination of the area and volume of discocytes (Ghosh, 1971;Leblond & Shoucri, 1978).…”

Section: Introductionmentioning

confidence: 99%

Effects of glutaraldehyde and critical point drying on the shape and size of erythrocytes in isotonic and hypotonic media

Eskelinen

Saukko

1983

Journal of Microscopy

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SUMMARY Scanning electron microscopy (SEM) was applied to studies on osmotically swollen erythrocytes fixed by a method using increasing concentrations of glutaraldehyde (GA), OsO4 as a post fixative, and critical point drying (CPD). This fixation method prevented the osmotic effect of GA itself and preserved the cellular equilibrium shape intact in various media, i.e. discocytes in an isotonic medium and ellipsoids or spheres and ghosts in a hypotonic medium. The discocytes were reduced less in diameter and area than the ellipsoids and spheres during CPD, but the degree of volume shrinkage was of the same magnitude (approx. 73%) regardless of the cell shape. The shrinkage of the discocytes was advanced until the dry volume (approx. 29 μm3) was reached, while the shrinkage of the osmotically swollen cells ceased earlier. A miniaturization process coupled with solvent evaporation is suggested as an explanation for this. The osmotic swelling of erythrocytes was studied quantitatively with the aid of SEM micrographs. The increase in volume was about 60%, which is in agreement with the values obtained by light microscopy for unfixed and GA‐fixed cells in the present work and with those reported in the literature.

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“…Mean particle size was calculated as outlined elsewhere (23). Apparent particle size using resistive pulse analysis is affected principally by the size of the particles, and by their shape and deformability (24). Since the shape of the red cell in the hypotonic solu-tions employed is close to spherical, no specific correction was made for the shape factor; aberration due to variable deformability was excluded by fixing the red cells prior to particle sizing in glutaraldehyde 0.125% at the osmolality used for including haemolysis.…”

Section: Methodsmentioning

confidence: 99%

“…(d). The peak of the volume/osmolality curve is taken as the maximum volume to which the cells can swell(22,24).The peaks of the curves obtained with hypotonic saline solutions containing 0.07, 2.2 and 4.4 mmol/l Ca2+ are shown inFig-ure 5: at a Ca2+ concentration of 2.2 mmol/l or greater the peak is significantly shifted to a lower osmolality than in saline alone. At 2.2 mmol/l Ca2+ the critical volume is unchanged, but it is slightly diminished at 4.4 mmol/l Ca2+.…”

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confidence: 99%

Abrogation of calcium exclusion by erythrocytes under hypotonic stress

Bowdler

Williams

Dougherty

1984

Scandinavian Journal of Haematology

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Normal human erythrocytes in isotonic suspension maintain a high gradient of calcium ion concentration across the membrane, with the cytosol [Ca2+] several orders of magnitude below plasma concentration. Despite the high degree of exclusion maintained against the entry of Ca2+, the osmotic resistance of red cells in hypotonic saline is significantly augmented by the presence in the medium of Ca2+in concentrations greater than 0.07 mmol/l. Resistive particle spectroscopy showed this to occur without alteration of the mean critical haemolytic volume; furthermore, in the presence of Ca2+there was a reduction in cell volume at prolytic osmolalities when compared with cells in calcium‐free sodium chloride solutions. This is shown to be due to augmentation of the prolytic loss of K+1in the presence of Ca2+ through calcium‐sensitive channels, and indicates that the pathway for passive K+efflux uncovered in the prolytic state is indistinguishable from that of normal passive K+diffusion. It is also demonstrated that the prolytic red cell membrane is permeable to Ca2+and admits the divalent cation to the inner aspect of the plasma membrane. These changes are comparable to those of cells prior to haemolysis in vivo, as in the irreversibly sickled cell, and are therefore not specific to the latter condition.

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Cell-membrane and rheological mechanisms: dynamic osmotic hemolysis of human erythrocytes and repair of ghosts, as studied by resistive pulse spectroscopy

Cited by 25 publications

References 0 publications

Alternative interpretation for the osmotic response of human erythrocytes

Alternative interpretation for the osmotic response of human erythrocytes

Effects of glutaraldehyde and critical point drying on the shape and size of erythrocytes in isotonic and hypotonic media

Abrogation of calcium exclusion by erythrocytes under hypotonic stress

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