Fast kinetic measurements and on-line dilution by flow injection cytometry.
An improved flow injection cytometry (FIC) system suitable for fast cellular kinetic measurements and on-line dilution is described. The instrument allows for measurements from 1.2 s (+/- 0.05 s) after the initiation of mixing, up to any time thereafter. Crucial factors in determining fast kinetic measurements, such as the displacement volume for sample introduction, rate of transport to cytometric interrogation point, and the mixing speed are evaluated and discussed. By varying the volume aspirated, this instrument can facilitate the dilution of cells and/or reagent over a range of one order of magnitude. The fast kinetic and on-line dilution capabilities were demonstrated by on-line staining of DNA in trout erythrocytes with 4',6-diamidino-2-phenylindole (DAPI). The utility of the instrument for measurement of enzyme kinetics was illustrated using human lymphocytes, measuring the glutathione S-transferase (GST) catalyzed reaction between monochlorobimane (MCB) and glutathione (GSH).
Lindberg
W
W
Department of Chemistry, University of Washington, Seattle 98195.
Scampavia
L D
LD
Ruzicka
J
J
Christian
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GD
D000818:Animals; D002021:Buffers; D002453:Cell Cycle; D004247:DNA / Q000032:analysis; D004867:Equipment Design; D004912:Erythrocytes / Q000648:ultrastructure; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D005978:Glutathione / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D007201:Indicator Dilution Techniques; D007202:Indicators and Reagents; D007211:Indoles; D007700:Kinetics; D008214:Lymphocytes / Q000201:enzymology; D009862:Online Systems; D012212:Rheology; D013048:Specimen Handling / Q000295:instrumentation*; D013194:Staining and Labeling; D014337:Trout
D002021:Buffers; D007202:Indicators and Reagents; D007211:Indoles; C007293:DAPI; D004247:DNA; D005982:Glutathione Transferase; D005978:Glutathione