Computer-assisted centrifugal elutriation. I. Detection system and data acquisition equipment.

To optimize cell separations by centrifugal elutriation, we constructed an on-line computer-controlled multiparametric light-scatter system. A bypass sample flow, at the outlet of an elutriation rotor, is hydrofocussed and three scatter parameters of each cell are determined up to a maximum of 15,000 cells/second. The 18-bit representation of the parameter values are cumulatively stored by means of a direct memory access interface. The histogram memory is continuously displayed to provide information on the number and type of cells that are elutriated. A special purpose operating system, implemented on a stand-alone computer configuration, allows a high data-acquisition rate and ample data processing capacity. In addition, a local network driver was constructed to facilitate off-line detailed analysis of the data. The equipment is well suited to monitor the centrigural elutriation process. The flexibility of the system allows an extension of the monitor to computer-controlled elutriation.

[1]  F. Bryant,et al.  Changes in total light scattering and absorption caused by changes in particle conformation. , 1968, Journal of theoretical biology.

[2]  R. Miller,et al.  The optimal application of forward and ninety-degree light scatter in flow cytometry for the gating of mononuclear cells. , 1985, Cytometry.

[3]  R. Weiner,et al.  Purification of human monocytes: isolation and collection of large numbers of peripheral blood monocytes. , 1980, Journal of immunological methods.

[4]  M. Meistrich.  Experimental Factors Involved in Separation by Centrifugal Elutriation , 1983 .

[5]  W. Grant,et al.  Resolution of cells by centrifugal elutriation. , 1979, Analytical biochemistry.

[6]  J. Brenman,et al.  Design principles for a counterflow centrifugation cell separation chamber. , 1976, Analytical biochemistry.

[7]  S. Pal,et al.  Polarization anisotropy in lidar multiple scattering from atmospheric clouds. , 1985, Applied optics.

[8]  M. Rapport,et al.  Slab gel analysis of the polypeptide components of rat brain subcellular organelles. , 1976, Analytical biochemistry.

[9]  J Swithenbank,et al.  Beam stop and vignetting effects in particle size measurements by laser diffraction. , 1986, Applied optics.

[10]  I T Young,et al.  A comparison of different focus functions for use in autofocus algorithms. , 1985, Cytometry.

[11]  C. Haanen,et al.  Monocyte purification with counterflow centrifugation monitored by continuous flow cytometry. , 1981, Journal of immunological methods.

[12]  D. Norris,et al.  Isolation of functional subsets of human peripheral blood monocytes. , 1979, Journal of immunology.

[13]  P. Sloot,et al.  Elastic light scattering from nucleated blood cells: rapid numerical analysis. , 1986, Applied optics.

[14]  J. Aubin.  Autofluorescence of viable cultured mammalian cells. , 1979, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[15]  R. S. Longhurst.  Geometrical and Physical Optics , 1967 .

[16]  R. A. Meyer,et al.  Light scattering from nucleated biological cells. , 1975, Biophysical journal.

[17]  P. Lindahl,et al.  On counter streaming centrifugation in the separation of cells and cell fragments. , 1956, Biochimica et biophysica acta.

[18]  H M Shapiro,et al.  Multistation multiparameter flow cytometry: a critical review and rationale. , 1983, Cytometry.

[19]  C. Figdor,et al.  Isolation of functionally different human monocytes by counterflow centrifugation elutriation. , 1982, Blood.

[20]  A. Fauci,et al.  An improved technique for the negative selection of large numbers of human lymphocytes and monocytes by counterflow centrifugation--elutriation. , 1980, Cellular immunology.

[21]  A. Donnenberg,et al.  Large scale separation of human bone marrow by counterflow centrifugation elutriation. , 1986, Journal of immunological methods.

[22]  M. Hartmann,et al.  Light scattering by small particles. Von H. C. VANDE HULST. New York: Dover Publications, Inc. 1981. Paperback, 470 S., 103 Abb. und 46 Tab., US $ 7.50 , 1984 .

[23]  C. Haanen,et al.  BONE MARROW REPOPULATION CAPACITY AFTER TRANSPLANTATION OF LYMPHOCYTE‐DEPLETED ALLOGENEIC BONE MARROW USING COUNTERFLOW CENTRIFUGATION , 1984, Transplantation.

[24]  H. V. Hulst.  Light Scattering by Small Particles , 1957 .

[25]  C. Figdor,et al.  Rapid isolation of mononuclear cells from buffy coats prepared by a new blood cell separator. , 1982, Journal of immunological methods.

[26]  R E Hand,et al.  Parametric analysis of histograms measured in flow cytometry. , 1983, Cytometry.

[27]  C. Figdor,et al.  Isolation of large numbers of highly purified lymphocytes and monocytes with a modified centrifugal elutriation technique. , 1981, Journal of immunological methods.

[28]  G C Salzman,et al.  Cell classification by laser light scattering: identification and separation of unstained leukocytes. , 1975, Acta cytologica.

[29]  C. Figdor,et al.  A centrifugal elutriation system of separating small numbers of cells. , 1984, Journal of immunological methods.

[30]  P. Lindahl,et al.  Principle of a Counter-streaming Centrifuge for the Separation of Particles of Different Sizes , 1948, Nature.

[31]  R. C. Benson,et al.  Cellular autofluorescence--is it due to flavins? , 1979, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[32]  Niklaus Wirth,et al.  Programming in Modula-2 , 1985, Texts and Monographs in Computer Science.

[33]  J Cuginin.  Selection and Use of General Purpose Programming Languages , 1984 .