Unified Flow Cytometer Setup
The calibration of flow cytometers in the clinical laboratory is of primordial importance for the interlaboratory comparison of quantitative and classification results.
Brightly fluorescent alignment beads are used to control the overall optical and electronic performance of a flow cytometer.
The measurement of a mixture of typically four fluorescent beads with known amounts of antibody binding sites or bound antibodies permits the calibration of the fluorescence scale of a flow cytometer in "bound fluorescent antibody molecules". A non fluorescent bead characterizes the assay noise level of the flow cytometer.
Besides the determination of the number of cell bound antibody molecules, the "sample window" of flow cytometers can be determined in absolute i.e. instrument independent terms (e.g the lower and upper threshold of displayed fluorescences may range from 200 to 2.000.000 molecules in a four logarithmic decade instrument).
Suitable calibration beads are available from different commercial sources. The characteristics and performance of various beads are displayed in the following chapters:
Becton Dickinson
USA:
Germany:
Flow Cytometry Standards Corporation (FCSC)
USA:
P.O.Box 194344
San Juan, PR 00919-4344
USA
Tel: +1/787/2741245
Fax: +1/787/7583267
E-mail: 73437.2722@compuserve.com
Europe:
FCS
Niels Bohrweg 11-13
P.O.Box 1336
NL-2302 BH Leiden
The Netherlands
Tel: +31/71/5236929
Fax: +31/71/5235594
E-mail: ebarel@POBOX.LeidenUniv.NL
Coulter/Immunotech
USA:
Germany:
Europark Fichtenhain B13
D-47807 Krefeld
Tel: +49/2151/333-5
Fax: +49/2151/333633
E-mail: --
Dako
Denmark:
Germany:
Am Stadtrand 52
D-22047 Hamburg
Tel: +49/40/696947-0
Fax: +49/40/6952741
E-mail: dako.gmbh@aol.com
Polysciences Inc.
USA:
400 Valley Road
Warrington, PA 18976
USA
Tel: +1/215/-343-6484
Fax: +1/215/-343-0214
E-mail: --
Germany:
Handelsstr. 3
D-69208 Eppelheim
Germany
Tel: +49/6221-76576
Fax: +49/6221-764620
E-mail: --
Sigma Biosciences
USA:
-- Immunochemicals Catalogue 1996 --
P.O.Box 14508
St. Louis, MO 63178-9916
Tel: +1/314-771-5750
Fax: +1/314-771-5757
Germany:
Grünwalder Weg 30
D-82041 Deisenhofen
Tel: +49/89/6513-0
Fax: +49/89/6513-1161
E-mail: --
Internet: Sigma-Aldrich, USA
Sigma-Aldrich, Germany
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