Richard M.P. Doornbos

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A method is described for measuring optical properties and deriving chromophore concentrations from diffuse reflection measurements at the surface of a turbid medium. The method uses a diffusion approximation model for the diffuse reflectance, in combination with models for the absorption and scattering coefficients. An optical fibre-based set-up, capable(More)
A flow cytometer using a solid state light source and detector was designed and built. For illumination of the sample stream two types of diode lasers (670 nm and 780 nm) were tested in a set-up designed to differentiate human leukocytes by means of light scattering. The detector is an avalanche photodiode, which was used to detect the weak scattered light(More)
This report describes a feasibility study concerning the use of a visible diode laser for two important fluorescence applications in a flow cytometer. With a 3 mW 635 nm diode laser, we performed immunofluorescence measurements using the fluorophore allophycocyanin (APC). We have measured CD8 positive lymphocytes with a two-step labeling procedure and the(More)
The complete scattering matrix S of spheres was measured with a flow cytometer. The experimental equipment allows simultaneous detection of two scattering-matrix elements for every sphere in the distribution. Two-parameter scatterplots with x and y coordinates determined by the S(ll) + S(ij) and S(ll)-S(ij) values are measured. Samples of spheres with very(More)
We investigated the fluorescence emission from three fluorophores commonly used for labeling cells in flow cytometry. We have demonstrated that the fluorescence emission from cells labeled with fluorescein-isothiocyanate (FITC), phycoerythrin (PE), and allophycocyanin (APC) is considerably saturated and bleached in standard flow cytometric conditions.(More)
We have developed an instrument for determination of the angular light scattering of beads and biological cells. The instrument uses radiation pressure for levitation of particles inside a cuvette. The setup consists of two 780-nm diode lasers in a vertical double-beam trapping configuration. In the horizontal direction a weakly focused 633-nm probe beam is(More)
Flow cytometric measurements of light scattering of polystyrene calibration beads revealed remarkable Lissajous-like loops in two-parameter scatter plots. The existence of such loops is shown to be in qualitative agreement with Lorenz-Mie scattering theory of homogeneous spheres. The occurrence of these patterns reflects the extreme particle size dependency(More)
Detailed analysis of elastic light scattering by spheres in FlowCytometers shows unexpected Lorenz-Mie scattering patterns. The complete scattering matrix S of spheres was measured. Two parameter scatterplots with xand ycoordinates determined by the S11 + Sij and S11 Sij values were obtained. Samples of spheres with very narrow size distributions were(More)
A computer program for graphical analysis of multidimensional flow cytometric list mode data is described. The program offers one-, two-, and three-dimensional inspection of an amount of data that is only limited by disk space. Subpopulations within the original data set can be identified by setting one or more two-dimensional AND gates around them. The(More)
We have constructed a simple device by which the optimal delay time between optical measurement of a cell and the application of the droplet charging pulse can be determined directly in a flow sorter. The device consists of a stainless steel chamber in which the sorted droplets are collected. In the collection chamber the collected droplets run through a(More)