Optical method and apparatus for red blood cell differentiation on a cell-by-cell basis, and simultaneous analysis of white blood cell differentiation

Methods and apparatus are disclosed for determining the volume, hemoglobin concentration, maturity and cell shape of mammalian red blood cells in a sample and simultaneously monitoring system standardization. Methods for distinguishing red blood cells from other cellular particles, prior to the red blood cell analysis are also disclosed. The method can be applied with accuracy over a wide range of visible spectrum. A whole blood sample is treated with a reagent solution containing a nonionic surfactant in an isotonic buffered solution at neutral pH, the red blood cells are passed through a beam of light in single file at a selected wavelength, obtaining an initial cytogram by means of the resultant magnitude of one light loss signal and one forward angle light scatter signal at a selected angular interval and a third side angle light scatter or two forward angle light scatter signals at selected angular intervals and a third side-angle light scatter signal, projecting the cytogram, point by point, onto a pre-calibrated 3-dimensional surface containing grid lines of volume and hemoglobin concentration, determining accurate values of cell volume and hemoglobin concentration by means of the location of each projected intercept onto the three dimensional grid surface. Methods and apparatus are disclosed for determining the volume, hemoglobin concentration, maturity and cell shape of mammalian red blood cells in a whole blood sample and simultaneously monitoring system standardization. Methods for distinguishing red blood cells from other cellular particles, prior to the red blood cell analysis are also disclosed. Red blood cells are passed through a beam of light in single file at a selected wavelength, obtaining an initial cytogram by means of the resultant magnitude of one light loss signal and one forward angle light scatter signal at a selected angular interval and a third side angle light scatter or two forward angle light scatter signals at a selected angular intervals and a third side-angle light scatter signal, projecting the cytogram, point by point, onto a pre-calibrated 3-dimensional surface containing grid lines of volume and hemoglobin concentration and determining accurate values of cell volume and hemoglobin concentration.