Cells were washed 3 times in cold PBS, mixed, harvested and lysed in Milli-Q water using a Cell Harvester; cell lysate was mixed with scintillation liquid (Optiphase Hisafe2, PerkinElmer, Shelton, CT) and 3H activity was measured in a scintillation counter (Tri-Carb 1600 TR, Packard, Meriden, CT).
The cellular DNA was collected on Whatman GFC filters using a harvester (Cell Harvester 8, Nunc), fixed with 96% ethanol, air dried and incorporated radioactivity was determined in the presence of 1 ml of scintillation solution (OptiPhos Hifase 3) in a liquid scintillation counter (Packard 1600 TR, Canberra Company).
[3H]Thymidine incorporation was determined on a liquid scintillation counter (Topcount, Perkin-Elmer) after harvesting (Tomtec cell harvester).
The plates were covered and incubated for 90 min. Finally, the cells were harvested using a cell harvester (Titertek Cell Harvester 550) and the quantity of bound I was expressed as picomoles/107 neutrophils.
[methyl-H]-Thymidine (20 μCi/mL) (PerkinElmer, Waltham, MA, USA) was added 24 hours before harvesting cells on fiberglass filters by means of an automated cell harvester (PerkinElmer).
Incorporated radioactivity was quantified by harvesting cells onto glass-fiber filters (Printer Filtermat A; Wallac, Turku, Finland) using an automatic cell harvester (Harvester 96 Mach III; Tomtec Inc, Orange, CT, USA) and then treated with melt-on scintillator sheets (MeltiLex A; Wallac) and counted with a scintillation beta counter (MicroBeta TriLux; Wallac).
Cultures were harvested using a cell harvester and thymidine incorporation was measured in a beta counter (Perkin Elmer).
The cells were incubated with 0.4 µCi of 3H-thymidine (specific activity: 20 Ci/mmol) for the last 18 hrs, trypsinized and harvested in a cell harvester.
Cells were harvested by a cell harvester (Inotech, Wohlen, Switzerland) on paper filters (Filtermat A, Wallac, Turku, Finland).
Cells were then rinsed with PBS, trypsinized, and harvested using a cell harvester (Cambridge Technology, Cambridge, MA, USA).
Plates were then frozen and defrosted, cells were harvested with a Cell Harvester, and cpm was measured with a 1205 BetaPlate liquid scintillation counter (Perkin Elmer, Waltham, MA, USA).
