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M Kerker

Publications and source records attributed to M Kerker.

7 recordsLinked to original sources
PubMed1983

Elastic and inelastic light scattering in flow cytometry.

A review of the fundamental aspects of elastic light scattering suggests that information about the shape and internal structure may best be obtained from signals measured in the backscattering directions. Size information can be most readily extracted from forward scattering signals. Spectral analysis of scattered signals with incident white light is a subject that merits further study. Signals from cells that have been stained with fluorescent dyes are proportional to dye content in the usual flow-cytometric configuration except in the case of dense, strongly anisometric structures such as sperm cells. The recent discovery that Raman signals from molecules adsorbed on small silver particles are strongly enhanced suggests the possibility of utilizing this effect for identification of molecular species inside biological cells.

Electromagnetic Fields↗
PubMed1982

Is the central dogma of flow cytometry true: that fluorescence intensity is proportional to cellular dye content?

Measurements and theoretical calculations of fluorescent emission from four samples of polystyrene microspheres (diameter 0.92, 1.63, 1.90 and 4.18 microns) containing the same fluorescent dye show a general dependence upon particle size, emission angle, and polarization conditions. However, for the excitation and detection conditions used in flow cytometry, the relative fluorescent intensities measured for the four particle sizes are proportional to the dye content to +10% accuracy, independent of particle size. Accordingly, the central dogma of flow cytometry 'that fluorescence is proportional to cellular dye content' is valid to this accuracy for these solid, highly refractive polymer particles. Most mammalian cells are much less refractive, therefore, should conform more closely to the central dogma.

Animals↗
PubMed1980

An optical model for fluorescence of mammalian sperm in flow cytometry.

When flat sperm heads that have been stained to fluorescence are examined in a flow cytometer, unexpectedly, skewed pulse height distributions are obtained despite the apparent homogeneity of the samples. This anomaly has been ascribed to an optical artifact that arises when the cells are oriented in flow. We have extended our model for fluorescent scattering to spheroids and here explore some aspects for oblate spheroids which serve to model sperm heads. Although computational limitations have restricted these studies to oblate spheroids about 1.5 micrometer in diameter and an eccentricity of 0.1, the results clearly show effects of particle size, shape, optical properties and particularly of orientation on the differential scattering cross-sections. This plethora of information contained in the fluorescent signals may suggest further experiments.

DNA↗
PubMed1979

Light scattering and fluorescence by small particles having internal structure.

We consider two related, yet distinct queries: 1. How does the internal morphology of a small particle affect the elastic light scattering signals? We have devised an algorithm, presently accurate for particles comparable only to small biological spheres (diameter less than 1 micron), which suggests that light scattering is sensitive to internal morphology only in the backward directions. Accordingly, observations should be obtained in these directions when probing for internal morphology. 2. How are fluorescent signals affected when the active molecules are variously distributed within small particles? One cannot assume that the fluorescent signals are simply proportional to the number of active molecules contained in the particle because there may also be a dependence upon the geometrical and optical properties of the particle and upon the particular spatial distribution of these molecules within the particle. Indeed, even the measured emission spectrum may be affected by such morphological features. Here, too, these calculations are mainly restricted to small particles (diameter less than 1 micron) in which the fluorescent molecules are isotropic and immobile. Under these conditions the effects are quite dramatic. These effects should be considered in quantitative procedures which utilize fluorescence for determining the concentration of specific molecules in small particles such as biological cells. They may provide a clue for discriminating among cells which differ morphologically or in which the spatial distribution of the fluorescent moiety differs. These effects may be minimized by utilizing a light source which is polarized perpendicularly to the scattering plane.

Cells↗