Normal human blood density gradient lymphocyte subset analysis: I. An interlaboratory flow cytometric comparison of 85 normal adults
✍ Scribed by Dr. G. E. Marti; L. Magruder; K. Patrick; M. Vail; W. Schuette; R. Keller; K. Muirhead; P. Horan; H. R. Gralnick
- Publisher
- John Wiley and Sons
- Year
- 1985
- Tongue
- English
- Weight
- 774 KB
- Volume
- 20
- Category
- Article
- ISSN
- 0361-8609
No coin nor oath required. For personal study only.
✦ Synopsis
An interlaboratory flow cytometric comparison of several commercially available human lymphocyte subset reagents was undertaken in three different laboratories. Fresh Hypaque-Ficoll purified blood mononuclear cells were stained at 4°C or 22°C. Direct or indirect surface immunofluorescence was carried out at all sites using an EPICS V flow cytometer. Fullbright, 10-pm fluorescent polystyrene microspheres were used for optical alignment and standardization. A log integral fluorescent histogram gated on forward and right angle scatter was collected on 1-2 X lo4 cells for each reagent and the proportion, of positive cell determined for each reagent. With the exception of one reagent, anti-B1, which showed an approximately twofold variation, all three laboratories showed remarkable agreement. Thus there was no significant difference noted for the following reagents: OKT4, CCM, Leu 3a, Leu 2a, OKT8, or CCTI. We attribute these findings to the availability of quality reagents, precision instrumentation, and a standard lymphocyte preparation.