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Biomedical subjects

R D Hiebert

Publications and source records attributed to R D Hiebert.

11 recordsLinked to original sources

Active analog pipeline delays for high signal rates in multistation flow cytometers.

Flow cytometers with sequential measuring stations require alignment of event timing information to assure that multiparameter data for each cell are properly correlated. Information from the first detector must be delayed until signals are produced at the last detector, with adjustments to align them in time for simultaneous digitization after the last measurement is completed. By using an analog "pipeline" delay, the deadtime between the first and last measuring stations can be minimized. The described device is capable of acquiring and propagating many signals within the delay period, with good fidelity at high signal rates. An integrated circuit charge-coupled device (CCD), which is an analog shift register, is shown to be useful as a signal delay in the time range from 22 microseconds to several milliseconds.

Flow Cytometry↗

Signal processing electronics for multiple electronic and optical measurements on cells.

Processing electronics for flow cytometry applications requiring simultaneous or sequential analysis of multiple electrical and optically sensed signals has been developed. A maximum of six analog signals are input to the processor. A measurement mode selector determines what signals are to be analyzed and the initial timing sequence of sense gates for acquiring signal crest values. Processor signal-triggering and sense gate time delays are selectable. Logic coincidence-anticoincidence circuits determine constraints on incoming signals. Gated peak-sense and hold signals are routed to computer interface electronics for digitizing and are then displayed as frequency distribution histograms using an LSI-11 computer. Signals also are processed as single parameters, ratios, and gated single parameters for output to a multichannel pulse-height analyzer and cell sorting electronics. The functional features of the processor are described along with examples illustrating simultaneous and sequential analysis of cultured cells stained with fluorescent dyes.

Animals↗

Modular electronics for flow cytometry and sorting: the LACEL system.

LACEL is newly developed, fast, general-purpose data acquisition and processing system for flow cytometric applications. The system's modular electronics allows flexibility in system configurations. The system can process as many as eight input analog parameters and can transfer 16-bit words between the user's electronics and the computer with standard input/output interfaces. The system's 8-fold coincidence logic capability can be set to operate with the noncoincidental timing that may occur in multiparameter flow measurements. As many as four parameters can be used to establish amplitude and timing criteria for each of two sorting directions. Two experiments can be on line with the computer at one time.

Cell Separation↗

A real-time delay monitor for flow-system cell sorters.

For optimum performance in cell sorting, it is critical to assure proper timing in the charging of droplets to be deflected. A method for determining the transiet delay time in cell sorters has been devised and applied to daily operation in the Los Alamos sorter systems. This delay monitor relies on detection of either scattered or absorbed light from cells in the fluid stream near the point of droplet breakoff.

Cell Separation↗

Pulse-height light-scatter distributions using flow-systems instrumentation.

Several laboratories have recently been making light-scatter measurements on cells and other particles using flow-systems instrumentation. We at the Los Alamos Scientific Laboratory, as well as others, have obtained multimodal pulse-height distributions in certain angular regimes from particles of supposedly uniform characteristics. Because it was assumed that multimodal distributions implied characteristics of multivalue, the accuracy of such data has been doubted. In the present work, pulse-height distributions anticipated on the basis of exact electromagnetic theory were calculated for particles of known characteristics. These calculated pulse-height distributions agree quite well with those obtained experimentally. Physical optics form the basis for the explanation of the complex pulse-height distributions obtained experimentally. However, the results of this study show that certain cautions are necessary in the interpretation of light-scatter data presented in this manner.

Autoanalysis↗

A flow-system multiangle light-scattering instrument for cell characterization.

A flow-system cell-analysis instrument is described in which cells from a heterogeneous population are characterized by their light-scatter patterns alone. As the cells pass at high speed through a focused helium/neon laser beam, the scatter pattern from each cell is sampled simultaneously at up to 32 angles between 0 degrees and 30 degrees with respect to the laser beam axis, and the scatter pattern for each cell is transferred to a computer. A mathematical clustering algorithm is used to determine the number of classes into which the cells can be divided, and a linear separation algorithm is used to find the boundaries between the classes. Preliminary results on exfoliated cells from gynecological specimens are presented. This technique may be useful for automated prescreening of gynecological specimens.

Autoanalysis↗