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United States Department of Agriculture

Agricultural Research Service

Title: Flow Cytometric Intracellular Ionized Calcium Analysis of Peripheral Blood Mononuclear Cells in Dairy Cattle with Fluo-4/fura Red Fluorescence Ratios

item Kimura, Kayoko - IOWA STATE UNIV., AMES
item Goff, Jesse
item Stabel, Judith
item Pesch, Bruce

Submitted to: Great Lakes International Imaging and Flow Cytometry Association
Publication Type: Abstract Only
Publication Acceptance Date: October 2, 1998
Publication Date: N/A

Technical Abstract: The measurement of changes in intracellular ionized calcium concentrations ([Ca2+]i) by flow cytometry is used to study responses of immune cells to activation of stimuli in humans and rodents. There are few applications for other animal species. We hope to establish a flow cytometric [Ca2+]i analysis method for PBMC and CD8-positive cells of cattle using Fluo-4 in combination with Fura-Red. Using anti-CD3 mAb (murine IgG1) and anti-muri to stimulate bovine PBMC, we were able to measure a 1.8 to 2.6 fold increase in [Ca2+]i in PBMC and a 2.2 to 3.2 fold increase in CD8 positive cells. The coefficient of variation was 4 to 10% for intra- and 4 to 18% for inter-assay variation, respectively. Although the analysis has certain limitation, we believe it will give us an alternative assay for immune cell function.

Last Modified: 4/22/2015
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