Sample handling for kinetics and molecular assembly in flow cytometry

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Description

Flow cytometry discriminates particle associated fluorescence from the fluorescence of the surrounding medium. It permits assemblies of macromolecular complexes on beads or cells to be detected in real-time with precision and specificity. The authors have investigated two types of robust sample handling systems which provide sub-second resolution and high throughput: (1) mixers which use stepper-motor driven syringes to initiate chemical reactions in msec time frames; and (2) flow injection controllers with valves and automated syringes used in chemical process control. In the former system, the authors used fast valves to overcome the disparity between mixing 100 {micro}ls of sample in ... continued below

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10 p.

Creation Information

Sklar, L. A.; Seamer, L. C.; Kuckuck, F.; Prossnitz, E.; Edwards, B. & Posner, G. July 1998.

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  • Los Alamos National Laboratory
    Publisher Info: Los Alamos National Lab., National Flow Cytometry Resource, NM (United States)
    Place of Publication: New Mexico

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Description

Flow cytometry discriminates particle associated fluorescence from the fluorescence of the surrounding medium. It permits assemblies of macromolecular complexes on beads or cells to be detected in real-time with precision and specificity. The authors have investigated two types of robust sample handling systems which provide sub-second resolution and high throughput: (1) mixers which use stepper-motor driven syringes to initiate chemical reactions in msec time frames; and (2) flow injection controllers with valves and automated syringes used in chemical process control. In the former system, the authors used fast valves to overcome the disparity between mixing 100 {micro}ls of sample in 100 msecs and delivering sample to a flow cytometer at 1 {micro}l/sec. Particles were detected within 100 msec after mixing, but turbulence was created which lasted for 1 sec after injection of the sample into the flow cytometer. They used optical criteria to discriminate particles which were out of alignment due to the turbulent flow. Complex sample handling protocols involving multiple mixing steps and sample dilution have also been achieved. With the latter system they were able to automate sample handling and delivery with intervals of a few seconds. The authors used a fluidic approach to defeat turbulence caused by sample introduction. By controlling both sheath and sample with individual syringes, the period of turbulence was reduced to {approximately} 200 msecs. Automated sample handling and sub-second resolution should permit broad analytical and diagnostic applications of flow cytometry.

Physical Description

10 p.

Notes

OSTI as DE98003496

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  • BIOS `98: an international symposium on biomedical optics, San Jose, CA (United States), 24-30 Jan 1998

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  • Other: DE98003496
  • Report No.: LA-UR--98-287
  • Report No.: CONF-980117--
  • Grant Number: W-7405-ENG-36
  • DOI: 10.2172/663512 | External Link
  • Office of Scientific & Technical Information Report Number: 663512
  • Archival Resource Key: ark:/67531/metadc703950

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Creation Date

  • July 1998

Added to The UNT Digital Library

  • Sept. 12, 2015, 6:31 a.m.

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  • Feb. 25, 2016, 4:42 p.m.

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Sklar, L. A.; Seamer, L. C.; Kuckuck, F.; Prossnitz, E.; Edwards, B. & Posner, G. Sample handling for kinetics and molecular assembly in flow cytometry, report, July 1998; New Mexico. (digital.library.unt.edu/ark:/67531/metadc703950/: accessed July 17, 2018), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.