Fcs gold

Patients with rapid progressive GCA were compared to a group of 12 age-matched patients retrospectively diagnosed with somatoform disorders without any signs of inflammatory CSF conditions (ie, <5 cells/μl CSF, <2 mmol/l lactate, no blood/CSF-barrier disruption, no intrathecal immunoglobulin synthesis [Reiber/OCB]). CSF samples were analyzed within 1 hour after lumbar puncture by centrifugation at 290 g for 15 minutes at 4 C in parallel with 100 μl peripheral blood. Blood-tinged CSF samples were excluded from the study. After treatment with VersaLyse buffer (Beckman Coulter) for 10 minutes, the samples were washed twice by addition of 3 ml FC-buffer (PBS [Sigma] supplemented with 2% heat-inactivated FCS Gold [BioSell] and 2 mM EDTA [Sigma]) and subsequent centrifugation at 290 g for 4 minutes. Following staining with CD14-FITC, CD3-PC5.5, CD56-PC7, CD4-APC, CD19-APC-AF700, CD8-PacificBlue, and CD45-KromeOrange (all Beckman Coulter), samples were washed once with FC buffer. After aspirating the supernatant, samples were resuspended and 20 μl flow count fluorospheres (Beckman Coulter) were added prior to acquisition using a Navios flow cytometer (Beckman Coulter). Resulting files were analyzed by Kaluza 2.1 (Beckman Coulter).