PubMed · 6135034
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1983-07-23. Block the phagocytes.. https://pubmed.ncbi.nlm.nih.gov/6135034/
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OBJECTIVES: Quantitation of Rh antibodies is important clinically in predicting the risk of hemolytic disease of the newborn. We describe a flow cytometry method for the quantitation of anti-D antibodies that we developed in parallel to a recently described method. METHODS: As a secondary antibody we used whole IgG instead of Fab molecules. The advantages, besides lower cost, include a strong fluorescence signal with no need for amplification, and the possibility of diluting samples to minimize the risk of agglutination by IgM antibodies. We did extensive studies on reproducibility. RESULTS: Reproducibility was superior to the autoanalyzer method. The two methods were roughly in agreement in estimating low, medium, or high levels of anti-D with a correlation coefficient of 0.89. The autoanalyzer measures the in vitro agglutination of all anti-D antibodies whereas flow cytometry measures the amount of IgG anti-D bound to red cells, which is more like the in vivo situation. CONCLUSION: Further studies in a clinical setting will show whether flow-cytometric quantitation may improve the diagnostic value of anti-D concentration measurement.
Simultaneous determination of the amounts of Na and K inside single human erythrocytes was accomplished by laser vaporization and monitoring the atomic emission produced. By using a modified sheath-flow arrangement, detection of 8 fg (0.35 fmol) of Na is possible with one laser pulse. By using Poisson statistics, one can obtain single-cell information even when multiple cells are vaporized per laser pulse. The intracellular contents for a given individual were found to vary significantly. The +/- 55% and +/- 155% variations for Na and K, respectively, cannot be explained by changes in cell volume. There is only a weak correlation between the Na and K contents in single cells. The results reflect the age distribution of erythrocytes in the sample. Presumably, the enzymes regulating ion transport lose their activities in the older cells.
Changes of plasma concentration are often used for in vitro characterisation of the hemolytic potency of artificial organs and apparatus. Different indices of hemolysis are derived from Hb concentration, which, in general, depend on experimental conditions and cannot be compared quantitatively or used to describe the in vivo damage. In this paper we propose a similarity number called "lysis number" that is independent of experimental conditions. It describes the probability for a single blood cell to be completely destroyed in a single pass through the corresponding artificial assist system. The concept is based on the steps: 1. Definition of "lysis number" as an index of hemolytic performance of artificial organs or implants. 2. Description of more complex hemolytic damaging processes (different hemolytic steps) that may be in series or parallel and definition of an effective lysis number. 3. Experimental in vitro estimation of each of the processes in consecutive steps. 4. Calculation of total hemolysis of the complex system using the linkage rules. 5. Application to in vivo by an appropriate differential equation in RBC mas taking into account mechanically-induced hemolysis rate, survival time of normal RBC and erythropoetic generation rate.