Sample handling for kinetics and molecular assembly in flow cytometry

Larry A. Sklar, Larry C. Seamer, Frederick Kuckuck, Richard G. Posner, Eric Prossnitz, Bruce Edwards, John P. Nolan

Research output: Chapter in Book/Report/Conference proceedingConference contribution

12 Citations (Scopus)

Abstract

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. We 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, we used fast valves to overcome the disparity between mixing 100 μls of sample in 100 msecs and delivering sample to a flow cytometer at 1μl/sec. Particles were detected within 100 msec after mixing, but unstable flow was created which lasted for 1 sec after injection of the sample into the flow cytometer. We used optical criteria to discriminate particles which were out of alignment due to the unstable flow. Complex sample handling protocols involving multiple mixing steps and sample dilution have also been achieved. With the latter system we were able to automate sample handling and delivery with intervals of a few seconds. We used a fluidic approach to defeat the instability caused by sample introduction. By controlling beth sheath and sample with individual syringes, the period of instability was reduced to ∼200 msecs. Automated sample handling and sub-second resolution should permit broad analytical and diagnostic applications of flow cytometry.

Original languageEnglish (US)
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
EditorsJ.R. Lakowicz, J.B. Alexander Ross
Pages144-153
Number of pages10
Volume3256
DOIs
StatePublished - 1998
Externally publishedYes
EventAdvances in Optical Biophysics - San Jose, CA, United States
Duration: Jan 25 1998Jan 26 1998

Other

OtherAdvances in Optical Biophysics
CountryUnited States
CitySan Jose, CA
Period1/25/981/26/98

Fingerprint

Syringes
cytometry
Flow cytometry
assembly
Kinetics
kinetics
Fluorescence
syringes
Fluidics
Dilution
Process control
Chemical reactions
Throughput
Controllers
injection
fluorescence
fluidics
sheaths
beads
assemblies

Keywords

  • Automation
  • Cell activation
  • Flow cytometry
  • Fluorescence
  • Kinetics
  • Ligand binding
  • Sample handling
  • Screening

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Condensed Matter Physics

Cite this

Sklar, L. A., Seamer, L. C., Kuckuck, F., Posner, R. G., Prossnitz, E., Edwards, B., & Nolan, J. P. (1998). Sample handling for kinetics and molecular assembly in flow cytometry. In J. R. Lakowicz, & J. B. Alexander Ross (Eds.), Proceedings of SPIE - The International Society for Optical Engineering (Vol. 3256, pp. 144-153) https://doi.org/10.1117/12.307057

Sample handling for kinetics and molecular assembly in flow cytometry. / Sklar, Larry A.; Seamer, Larry C.; Kuckuck, Frederick; Posner, Richard G.; Prossnitz, Eric; Edwards, Bruce; Nolan, John P.

Proceedings of SPIE - The International Society for Optical Engineering. ed. / J.R. Lakowicz; J.B. Alexander Ross. Vol. 3256 1998. p. 144-153.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Sklar, LA, Seamer, LC, Kuckuck, F, Posner, RG, Prossnitz, E, Edwards, B & Nolan, JP 1998, Sample handling for kinetics and molecular assembly in flow cytometry. in JR Lakowicz & JB Alexander Ross (eds), Proceedings of SPIE - The International Society for Optical Engineering. vol. 3256, pp. 144-153, Advances in Optical Biophysics, San Jose, CA, United States, 1/25/98. https://doi.org/10.1117/12.307057
Sklar LA, Seamer LC, Kuckuck F, Posner RG, Prossnitz E, Edwards B et al. Sample handling for kinetics and molecular assembly in flow cytometry. In Lakowicz JR, Alexander Ross JB, editors, Proceedings of SPIE - The International Society for Optical Engineering. Vol. 3256. 1998. p. 144-153 https://doi.org/10.1117/12.307057
Sklar, Larry A. ; Seamer, Larry C. ; Kuckuck, Frederick ; Posner, Richard G. ; Prossnitz, Eric ; Edwards, Bruce ; Nolan, John P. / Sample handling for kinetics and molecular assembly in flow cytometry. Proceedings of SPIE - The International Society for Optical Engineering. editor / J.R. Lakowicz ; J.B. Alexander Ross. Vol. 3256 1998. pp. 144-153
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