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Proceedings Paper

Quantification of chemotaxis during pediatric cardiac surgery by flow and laser scanning cytometry
Author(s): Attila Tarnok; Joerg W. Schmid; Pavel Osmancik; Dominik Lenz; Michal Pipek; Joerg Hambsch; Andreas O.H. Gerstner; Peter Schneider
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Paper Abstract

Cardiac surgery with cardiopulmonary bypass (CPB) alters the leukocyte composition of the peripheral blood (PB). This response contributes to the sometimes adverse outcome with capillary leakage. Migration of activated cells to sites of inflammation, driven by chemokines is part of this response. In order to determine the chemotactic activity of patients serum during and after surgery we established an assay for PB leukocytes (PBL). PBL from healthy donors were isolated and 250,000 cells were placed into a migration chamber separated by a filter from a second lower chamber filled with patient serum. After incubation cells from top and bottom chamber were removed and stained with a cocktail of monoclonal antibodies for leukocyte subsets and analyzed on a flow cytometer (FCM). Cells at the bottom of the filter belong to the migrating compartment and were quantified by LSC after staining of nucleated cells. Increased chemotactic activity started at onset of anaesthesia followed by a phase of low activity immediately after surgery and a second phase of a high post-operative activity. The in vitro results correlated with results obtained by immunopenotyping of circulating PBL. Manipulation of the chemokine pattern might prove beneficial to prevent extravasation of cells leading to tissue damage. In chemotaxis assays with low amount of available serum the combined use of FCM and Laser Scanning LSC proved as an appropriate analytical tool.

Paper Details

Date Published: 28 May 2002
PDF: 11 pages
Proc. SPIE 4622, Optical Diagnostics of Living Cells V, (28 May 2002); doi: 10.1117/12.468363
Show Author Affiliations
Attila Tarnok, Univ. Leipzig (Germany)
Joerg W. Schmid, Univ. Leipzig (Germany)
Pavel Osmancik, Univ. Hospital Kralovske Vinohrady/Charles Univ. (Germany)
Dominik Lenz, Univ. Leipzig (Germany)
Michal Pipek, Univ. Leipzig (Germany)
Joerg Hambsch, Univ. Leipzig (Germany)
Andreas O.H. Gerstner, Univ. Leipzig (Germany)
Peter Schneider, Univ. Leipzig (Germany)

Published in SPIE Proceedings Vol. 4622:
Optical Diagnostics of Living Cells V
Robert C. Leif; Daniel L. Farkas; Robert C. Leif, Editor(s)

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