The purpose of this research is to develop automated sample and solution handling procedures in the analysis of cells, cell organelles and chemical substances of relevance in that area. In this work the principles and methodology of flow injection analysis (FIA) will be adopted and further developed for that purpose. For detection, flow fluorimetry and flow cytometry will be used. The developed technology will allow kinetic analysis of faster cell reactions than is possible with present methodology. Further, using FIA, accurate temporal, stoichiometric and temperature control in reagent-cell and agonist/antagonist-cell reactions becomes available. A third area of research is automated separation and analysis of cells and cell related substances. This field will be pursued using in-line filters in combination with flow direction reversals as well as antibody labeled magnetic particles controlled by electromagnets. The precise timing of experimental events such as valve switching, setting flow speeds, flow stop/go, flow reversals, electromagnetic capturing and so on, will be done using computer (PC) controlled instruments.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM045260-02
Application #
3304661
Study Section
Special Emphasis Panel (SSS (C))
Project Start
1991-01-01
Project End
1993-12-31
Budget Start
1992-01-01
Budget End
1992-12-31
Support Year
2
Fiscal Year
1992
Total Cost
Indirect Cost
Name
University of Washington
Department
Type
Schools of Arts and Sciences
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195
Wu, Chao-Hsiang; Scampavia, Louis; Ruzicka, Jaromir (2002) Microsequential injection: anion separations using 'lab-on-valve' coupled with capillary electrophoresis. Analyst 127:898-905
Carroll, Andrea D; Scampavia, Louis; Ruzicka, Jaromir (2002) Label dilution method: a novel tool for bioligand interaction studies using bead injection in the lab-on-valve format. Analyst 127:1228-32
Schulz, Craig M; Scampavia, Louis; Ruzicka, Jaromir (2002) Real-time monitoring of lactate extrusion and glucose consumption of cultured cells using a lab-on-valve system. Analyst 127:1583-8
Schulz, Craig M; Ruzicka, Jaromir (2002) Real-time determination of glucose consumption by live cells using a lab-on-valve system with an integrated microbioreactor. Analyst 127:1293-8
Lahdesmaki, I; Ruzicka, J; Ivaska, A (2000) Novel flow injection methods for drug-receptor interaction studies, based on probing cell metabolism. Analyst 125:1889-95
Hodder, P S; Beeson, C; Ruzicka, J (2000) Equilibrium and kinetic measurements of muscarinic receptor antagonism on living cells using bead injection spectroscopy. Anal Chem 72:3109-15
Lahdesmaki, I; Scampavia, L D; Beeson, C et al. (1999) Detection of oxygen consumption of cultured adherent cells by bead injection spectroscopy. Anal Chem 71:5248-52
Scampavia, L D; Hodder, P S; Lahdesmaki, I et al. (1999) Automation of functional assays by flow injection fluorescence microscopy. Trends Biotechnol 17:443-7
Connors, W L; Ruzicka, J (1999) Flow injection microscopy for the study of intracellular calcium mobilization by muscarinic agonists. Anal Biochem 268:377-82
Hodder, P S; Ruzicka, J (1999) A flow injection renewable surface technique for cell-based drug discovery functional assays. Anal Chem 71:1160-6

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