The specific aims are to develop novel techniques and devices for experimentation on cells as performed in flow cytometry and microscopy. This project will be based on flow injection analysis, which is a well established technique for the automated, fluidic handling of samples and reagents in analytical chemistry. Central to this endeavor is that greater sophistication in fluid handling will enhance current research in cell biology and drug discovery by providing reliable and reproducible measurements. Moreover, unique information on the kinetics of biologcial processes in time frames and experimental designs inaccessible by any other means can be obtained. The long term objectives are the development of: (1) rapid mixing devices/protocols for kinetic analyses in flow cytometry, (2) fluidic based cell manipulation and sorting for imaging cytometry using microfabricated flow chambers, (3) cell stimulation methods for work in drug discovery and fundamental stimulus/receptor interactions and (4) novel perifusion devices for the measurements of cellular secretion products with respect to intracellular signals. The results of the proposed work will provide researchers with new methods for experimentation and cell handling in fields such as pharmacology, pathology, immunology, hematology and the biotechnological industry. Benefits will impact basic resaerch involved in elucidating dynamic or kinetic cellular interactions. Moreover, practical research that requires automated methods for testing/evaluating cells with respect to drug/agonist, antibody and protein synthesis will also benefit.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM045260-10S1
Application #
6418610
Study Section
Special Emphasis Panel (ZRG2 (37))
Program Officer
Lewis, Catherine D
Project Start
1991-01-01
Project End
2001-12-31
Budget Start
2000-01-01
Budget End
2001-12-31
Support Year
10
Fiscal Year
2001
Total Cost
$77,047
Indirect Cost
Name
University of Washington
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
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
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
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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