[3H] incorporation was determined using a TopCount scintillation counter (Packard Instrument).
Cells were then harvested and the H-thymidine incorporation was measured using a Topcount NXT counter (PerkinElmer, Waltham, MA, USA).
Migrating cells were detected by the use of CellTiter Glo® Dye (Promega, Southampton, UK) and resulting luminescence measured using a TopCount scintillation counter (Perkin Elmer, Waltham, MA, USA).
Migrating cells were detected by the use of CellTiter Glo® Dye (Promega, Southampton, UK) and resulting luminescence measured using a TopCount scintillation counter (PerkinElmer, Waltham, MA, USA).
The cells were then harvested onto a Packard Unifilter Plate using a MicroMate 196 Cell Harvester and counted using a TopCount Microplate Scintillation Counter (Packard Instruments, Meriden, CT).
3H-labeled thymidine incorporation was measured using a Topcount liquid scintillation counter (Packard Bioscience) and results expressed as stimulation indices (S.I ., defined as the ratio of the mean counts per minute (cpm) of wells incubated with HA antigen to the mean cpm of wells incubated with medium alone.
Plates were then sealed, and the signal was left to stabilise for 1 h in the dark before luminescence was determined using a TopCount NXT HTS counter (PerkinElmer).
During the last 24 h, cultures were pulsed with 1 μCi/well [H]TdR, and uptake was detected using a Topcount microplate scintillation counter and results are expressed as mean of triplicate cultures.
Cellular proliferation was detected by 3H-thymidine incorporation into replicating DNA was measured by adding 1 µCi to each replicate well of 105 PBMC for 24 hours before quantitative analysis using a Topcount® Liquid Scintillation Counter (Perkin-Elmer, Waltham, MA).
The cells were harvested, and [H]thymidine uptake was determined using a TopCount NXT microplate scintillation counter.
