scholarly journals MEASUREMENT OF PROTEIN CONCENTRATION BY QUANTITATIVE ELECTRON MICROSCOPY

1969 ◽  
Vol 40 (3) ◽  
pp. 773-778 ◽  
Author(s):  
Lloyd Silverman ◽  
David Glick
Keyword(s):  
Electron Microscopy ◽  
Protein Concentration ◽  
Clinical Laboratory ◽  
Red Cells ◽  
Cell Protein ◽  
Red Cell ◽  
Protein Loss ◽  
Quantitative Electron Microscopy ◽  
Thin Sections ◽  
Mean Values

The method of quantitative electron microscopy was applied to the measurement of protein concentration in thin sections. The human erythrocyte was selected as a model because of its apparently uniform protein concentration. Phosphotungstic acid (PTA) in aqueous solution was used as a reversible stain for protein, and PTA-stained Dowex resin spheres were embedded along with the red cells as standards for measurement of section thickness. The mass of stain removed from a given area of sectioned red cell by buffer (pH 7.4) was measured by quantitative electron microscopy. From the stoichiometry of the reaction between PTA and red cell protein established in this study, the amount of protein present in the measured area was calculated. From this amount of protein and the measured thickness, the concentration of protein was calculated and expressed as g/100 ml, for comparison with the clinical laboratory value for hemoglobin. Groups of red cells from the same sample were measured on 3 different days and their mean values (g/100 ml ± SD) were 29 ± 3.9, 30 ± 2.7, and 33 ± 4.6, compared to the clinical laboratory value of 32.1 g/100 ml packed cells, after correction for volume change and protein loss during fixation.

scholarly journals The Relationship of Bartonella Bacilliformis to the Red Blood Cell as Revealed by Electron Microscopy

Blood ◽  
1969 ◽  
Vol 33 (5) ◽  
pp. 708-716 ◽  
Author(s):  
MANUEL CUADRA ◽  
JUAN TAKANO
Keyword(s):  
Electron Microscopy ◽  
Blood Cell ◽  
Red Cells ◽  
Red Cell ◽  
Bartonella Bacilliformis ◽  
Coccoid Form ◽  
Blood Smears ◽  
Ultrathin Sections ◽  
Relationship Of ◽  
The Relationship

Abstract Ultrathin sections of erythrocytosis parasitized by B. bacilliformis have been examined by electron microscopy. The study concerns three Oroya Fever patients whose blood smears showed B. bacilliformis predominantly in its coccoid form as parastizing over 70 per cent of the red cells. B. bacilliformis is termed as a bacterium in its structure and appears to lie not only on the host red cells but predominantly within them. Therefore, this organism might have the capacity to penetrate into the red cell. This finding does not change the basic concept regarding the mechanism of the anemia of Oroya Fever.

Scanning Electron Microscopy of Elliptocytes in Man and Llama

1975 ◽  
Vol 33 ◽  
pp. 510-511
Author(s):  
Matias Pardo ◽  
Robert C. Grauer ◽  
James H. Swart ◽  
Robert J. Hartsock
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Electron Microscope ◽  
Autosomal Dominant ◽  
Central Zone ◽  
Red Cells ◽  
Red Cell ◽  
Shadow Casting ◽  
Dominant Disease ◽  
Scanning Electron

Elliptical red cells may be found in man, camels, llamas, birds and reptiles. In man, these cells can occur as an autosomal dominant disease. Using a shadow-casting technic, Rebuck and Van Slyck found a constant bipolar massing of hemoglobin in the elliptical cells of man. This polar massing of hemoglobin was not observed in the red cells of the llama. From these observations they deduced that the elliptical cells in man were biconcave whereas the cells from the llama were uniformly elliptical without a zone of central thinning.The scanning electron microscope was used to confirm these observations by comparing the red cells from a patient with hereditary elliptocytosis with red cells from a llama. As shown in Fig. 1, the elliptical cells of man are canoe-shaped. Bipolar massing of hemoglobin in these cells is seen in the red cell in Fig. 3. These observations are contrasted with red cells from the llama which are also elliptical, but they lack a central zone of thinning as shown in Fig. 2.

scholarly journals MEASUREMENT OF THICKNESS WITHIN SECTIONS BY QUANTITATIVE ELECTRON MICROSCOPY

1969 ◽  
Vol 40 (3) ◽  
pp. 768-772 ◽  
Author(s):  
Lloyd Silverman ◽  
Berit Schreiner ◽  
David Glick
Keyword(s):  
Electron Microscopy ◽  
Phosphotungstic Acid ◽  
Exchange Resin ◽  
Interference Microscopy ◽  
Quantitative Electron Microscopy ◽  
Thin Sections ◽  
Calculated Mass ◽  
Thickness Standard ◽  
Embedding Medium ◽  
Measurement Of Thickness

To apply the method of quantitative electron microscopy to the measurement of mass in thin sections, the thickness of the section at or very near the structure to be studied must be known. Dowex anion exchange resin AG 1 x 2, stained with phosphotungstic acid (PTA) at pH 6.4, was used as a thickness standard which could be embedded and sectioned. The sectioned PTA-Dowex appeared uniformly stained and exhibited suitable electron opacity. The stoichiometry of the reaction between PTA and the Dowex resin was measured by three independent methods based on gravimetric, colorimetric, and nitrogen determinations whose results showed close agreement. From the PTA uptake, the density of the stained spheres was calculated. Mass of a defined area of PTA-Dowex was measured by quantitative electron microscopy, and from this mass and density, the volume and then the thickness were calculated. The values for thickness were compared to those obtained by interference microscopy on the embedding medium alone in the same sections.

Foaming and Emulsifying Properties of Porcine Red Cell Protein Concentrate

2010 ◽  
Vol 16 (4) ◽  
pp. 289-296 ◽  
Author(s):  
P. Salvador ◽  
E. Saguer ◽  
D. Parés ◽  
C. Carretero ◽  
M. Toldrà
Keyword(s):  
Protein Content ◽  
Spray Drying ◽  
Protein Concentration ◽  
Cell Protein ◽  
Blood Protein ◽  
Red Cell ◽  
Emulsifying Properties ◽  
Food Ingredient ◽  
Foaming Capacity ◽  
Spray Dried

This work focuses on studying the effects of pH (7.0 and 4.5) and protein concentration on the foaming and emulsifying properties of fresh (F) and spray-dried (SD) porcine red cell protein (RCP) concentrates in order to evaluate the proper use of this blood protein as a functional food ingredient. Also, protein solubility is measured through the pH range from 3.0 to 8.0. In each case, all concentrates show a high solubility, although this is significantly affected by pH. Spray drying slightly reduces the solubility at mild acid and neutral conditions. The foaming capacity is found to be dependent on pH as well as on the drying treatment. SD-RCP concentrates show better foaming capacity than F-RCP. The minimum protein concentration required to attain the highest foaming capacity is found under acid pH for the spray-dried concentrates. Although F-RCP shows low foam stability at acid and neutral pH, spray drying and protein content enhance the stability of foams. Emulsifying properties show dependence on pH as well as on protein content. Furthermore, spray drying affects the emulsifying properties but in different ways, depending on pH and protein concentration.

Fibrinogen-I131, T-1824, and red cell-Cr51 spaces following hemorrhage

1963 ◽  
Vol 205 (3) ◽  
pp. 527-532 ◽  
Author(s):  
Carleton H. Baker
Keyword(s):  
Plasma Protein ◽  
Protein Concentration ◽  
Mixing Time ◽  
Red Cells ◽  
Red Cell ◽  
Plasma Protein Concentration ◽  
Volume Determination ◽  
Indicator Concentration ◽  
Fluid Shifts ◽  
Concentration Changes

Altered hemodynamics following hemorrhage causing extended mixing time of indicators, fluid shifts, and changes in indicator disappearance slopes were studied in 17 anesthetized splenectomized dogs. Fibrinogen-I131, T-1824, and red cell-Cr51 spaces were determined simultaneously and the animals were bled 11.3 ml/kg. There was a significant decline in concentration of the three indicators that could not be accounted for on the basis of hematocrit or plasma protein concentration changes. After 20 min stabilization, the spaces were again measured, and the animals were bled an average of 17.7 ml/kg (to an arterial pressure of 60 mm Hg). The decline in indicator concentration was again observed. After allowing the animals to stabilize for 20 min, the spaces were again measured. There was a mobilization of red cells following the first hemorrhage and a trapping of red cells following the second hemorrhage. Fibrinogen-I131 and T-1824 spaces agreed closely with the expected spaces calculated from changes in hematocrit or plasma protein concentration. The fibrinogen space was consistently less than the T-1824 space. The ratio BVcells/ BVfibrinogen significantly increased from 0.92 to 0.99 following the hemorrhages. This suggested a possible redistribution of "extra plasma" or a smaller involvement of extravascular space in the plasma volume determination. BVcells/ BVT-1824 did not change following the hemorrhages. Possible causes for the decline in indicator concentration following hemorrhage are discussed.

scholarly journals Effect of phosphatidylserine on the shape of McLeod red cell acanthocytes

Blood ◽  
1989 ◽  
Vol 74 (5) ◽  
pp. 1826-1835 ◽  
Author(s):  
CM Redman ◽  
T Huima ◽  
E Robbins ◽  
S Lee ◽  
WL Marsh
Keyword(s):  
Electron Microscopy ◽  
Red Cells ◽  
Cytoskeletal Protein ◽  
Red Cell ◽  
Enzymatic Process ◽  
Membrane Bilayer ◽  
Irregular Surfaces ◽  
The Common ◽  
Shape Changes ◽  
Scanning Electron

Abstract The rare McLeod blood group phenotype is characterized by weak Kell antigens, lack of the common Kx antigen, and acanthocytic morphology. Previous studies that did not detect membrane or cytoskeletal protein abnormalities suggested a lipid disturbance. In normal red cells, dimyristoyl phosphatidylserine (DMPS) is transported across the membrane by an enzymatic process and accumulates in the inner leaflet of the membrane bilayer causing discocyte to stomatocyte shape changes. Scanning electron microscopy of McLeod red cells shows a mixture comprised of 15% discocytes, 51% with irregular surfaces, and 34% acanthocytes. On incubation with various concentrations of DMPS at 37 degrees C for periods up to two hours, McLeod red cells transported DMPS across the membrane and caused irregularly shaped and acanthocytic McLeod red cells to attain normal discocyte shape and later to become stomatocytes. Chlorpromazine, which at 0 degrees C preferentially partitions into the inner monolayer of the membrane, had a similar effect on the shape of McLeod red cells. This suggests that in McLeod cells acanthocytosis is due to a lack of lipid in the inner leaflet of the membrane bilayer but that the imbalance is not caused by defective transport of phosphatidylserine across the membrane.

scholarly journals Electron Microscopy of the Red Cells in Erythropoietic Porphyria

Blood ◽  
1965 ◽  
Vol 25 (1) ◽  
pp. 49-55 ◽  
Author(s):  
SAMUEL GROSS ◽  
MELVIN D. SCHOENBERG ◽  
VIRGIL R. MUMAW
Keyword(s):  
Electron Microscopy ◽  
Cell Lines ◽  
Red Cells ◽  
Red Cell ◽  
Heme Synthesis ◽  
Cell Series

Abstract With the aid of electron microscopy two different red cell lines have been identified in erythropoietic porphyria. A normal red cell series has been found in association with hemoglobin containing normoblastic nuclei and ferritin laden reticulocytes. The abnormal line presumably represents the porphyrin containing cells. A possible explanation to account for the abnormality in heme synthesis has been proposed.

scholarly journals STUDIES IN THE BLOOD CYTOLOGY OF THE RABBIT

1930 ◽  
Vol 52 (1) ◽  
pp. 23-38 ◽  
Author(s):  
Louise Pearce ◽  
Albert E. Casey
Keyword(s):  
Peripheral Blood ◽  
Narrow Range ◽  
Red Cells ◽  
Red Cell ◽  
Hemoglobin Content ◽  
Individual Group ◽  
Mean Values ◽  
Range Of Variation

Observations are reported on the consecutive weekly erythrocyte counts and the hemoglobin contents of the peripheral blood in 5 groups of normal rabbits, comprising 45 animals, during a period of 20 months from October, 1927 to July, 1929. The duration of individual group examinations varied from 8 to 35 weeks. The results are analyzed on the basis of the weekly mean values of each group. On the whole, the erythrocyte values were quite uniform within a narrow range of variation, while the hemoglobin content was comparatively irregular within a wider range of variation. The major changes in the levels of mean values of both the red cells and the hemoglobin, however, were found to be statistically significant. The directions or trends in the levels of the erythrocyte and hemoglobin mean values did not necessarily move in opposite directions. The general levels of the erythrocyte and hemoglobin mean values were not identical for two consecutive years, those of 1927–28 being higher than those of 1928–29. The fluctuations of both red cell and hemoglobin mean values observed in one group of animals were also usually observed in another group examined during the same months.

Blood reservoirs in the splenectomized dog

1965 ◽  
Vol 208 (3) ◽  
pp. 485-491 ◽  
Author(s):  
Carleton H. Baker
Keyword(s):  
Arterial Pressure ◽  
Total Peripheral Resistance ◽  
Protein Concentration ◽  
Peripheral Resistance ◽  
Red Cells ◽  
Red Cell ◽  
Epinephrine Infusion ◽  
Plasma Protein Concentration ◽  
Percent Change ◽  
Cold Pack

Changes in plasma protein concentration, hematocrit, and red cells Cr51 concentration (count/min per ml blood) were determined following various procedures to acutely alter hemodynamics in an attempt to determine the presence of red cell or plasma reservoirs in splenectomized dogs. The cell reservoir index (percent change in hematocrit minus the percent change in count per minute per milliliter blood) increased significantly following procedures causing increased total peripheral resistance and arterial pressure. These procedures included epinephrine, angiotensin, and norepinephrine infusions as well as cold-pack applications. This was interpreted to be a shift of red cells out of reservoirs. Depressor procedures such as phentolamine followed by epinephrine infusion, isoproterenol, and acetylcholine caused no change in reservoir index. Histamine infusion caused a reduction in cell reservoir index. There was a shift of whole plasma from reservoirs into the active circulation following the infusion of epinephrine into animals previously administered phentolamine. The red cell reservoirs are believed to be generally distributed throughout the organism.

Effect of phosphatidylserine on the shape of McLeod red cell acanthocytes

Blood ◽  
1989 ◽  
Vol 74 (5) ◽  
pp. 1826-1835
Author(s):  
CM Redman ◽  
T Huima ◽  
E Robbins ◽  
S Lee ◽  
WL Marsh
Keyword(s):  
Electron Microscopy ◽  
Red Cells ◽  
Cytoskeletal Protein ◽  
Red Cell ◽  
Enzymatic Process ◽  
Membrane Bilayer ◽  
Irregular Surfaces ◽  
The Common ◽  
Shape Changes ◽  
Scanning Electron

The rare McLeod blood group phenotype is characterized by weak Kell antigens, lack of the common Kx antigen, and acanthocytic morphology. Previous studies that did not detect membrane or cytoskeletal protein abnormalities suggested a lipid disturbance. In normal red cells, dimyristoyl phosphatidylserine (DMPS) is transported across the membrane by an enzymatic process and accumulates in the inner leaflet of the membrane bilayer causing discocyte to stomatocyte shape changes. Scanning electron microscopy of McLeod red cells shows a mixture comprised of 15% discocytes, 51% with irregular surfaces, and 34% acanthocytes. On incubation with various concentrations of DMPS at 37 degrees C for periods up to two hours, McLeod red cells transported DMPS across the membrane and caused irregularly shaped and acanthocytic McLeod red cells to attain normal discocyte shape and later to become stomatocytes. Chlorpromazine, which at 0 degrees C preferentially partitions into the inner monolayer of the membrane, had a similar effect on the shape of McLeod red cells. This suggests that in McLeod cells acanthocytosis is due to a lack of lipid in the inner leaflet of the membrane bilayer but that the imbalance is not caused by defective transport of phosphatidylserine across the membrane.

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