BD FACSort Manuals

1 BD FACSort System 337452 Rev. A BD FACSort System After completing this module, you will be able to: • Describe the BD FACSort™ system and how it works. • Perform instrument startup and shutdown procedures. • Describe instrument maintenance procedures. 2 BD FACSort System 337452 Rev. A BD FACSort System The BD FACSort™ system is an automated flow cytometer and cell sorter. It analyzes cells as they pass through a focused laser beam one at a time in a moving fluid stream. A BD FACSort flow cytometer can be used for routine research applications, immunophenotyping, and DNA cell-cycle analysis. In addition, the BD FACSort flow cytometer allows you to identify and sort (isolate) a subpopulation from your sample. The BD FACSort system consists of a sensor module (Figure 3-1), computer module, and several software packages. Many of the cytometer functions are controlled by the software. Cytometer Figure 3-1 BD FACSort Sensor Module Power Switch This switch, located on the upper left corner of the instrument, turns the instrument on and off. 3 BD FACSort System 337452 Rev. A Fluidics Control Panel This panel consists of the fluidics control dial and the sample flow rate buttons. Fluidics Control Dial This rotary dial allows you to select from five fluidic modes. DRAIN —forces liquids out of the flow cell and fluid lines leading to the waste. FILL —fills the flow cell and fluid lines with sheath fluid at a controlled rate, which prevents bubble formation. RUN —pressurizes the sample tube to transport the cell sample suspension through the sample injection tube and into the flow cell. When a tube is not on the Sample Injection Port (SIP), the cytometer goes into auto Standby. Sheath flow is restricted and laser power is lowered. STANDBY —stops sheath flow and lowers laser power to conserve sheath fluid and prolong laser life • Hard STANDBY —When the fluid control dial is manually turned to STANDBY, all fluid valves are closed except for one valve that vents the sample test tube air pressure to the atmosphere. Additionally, the laser input current is reduced to a standby level. • Auto STANDBY —If the instrument is in RUN and the sample test tube is removed, the instrument goes into autostandby. Twelve seconds later, the system shuts off the sheath flow by means of a pinch valve between the flow cell and the waste reservoir. Additionally, the laser input current is reduced to a standby level. In this mode, back pressure is maintained to flush the sample injection tube. The system returns to RUN mode when you reinstall the sample tube. The pinch valve is opened and the laser returns to full power. lollo LO MED HI 4 BD FACSort System 337452 Rev. A BACKFLUSH —reverses the flow of sheath fluid and flushes fluid out of the sample injection tube to remove clogs. Sample Flow Rate Buttons Three buttons allow you to select the sample flow rate. • LO —12 µ L/min of sample through the flow cell • MED —35 µ L/min of sample through the flow cell • HI —60 µ L/min of sample through the flow cell Fluidics Drawer The fluidics drawer (the lower left panel of the instrument), slides out for easy access to the fluidics reservoirs and sheath filter. Sheath Reservoir This 4-liter container, located on the left, holds enough sheath fluid for approximately 3 hours of run time. It is equipped with a low-level fluid detector which indicates via software a near-empty condition. A metal bracket fits over the reservoir to secure the tank. VENT VALVE PRESS TO RELIEVE PRESSURE SALINE FILTER J69 J70 WASTE SHEATH sheath tank waste tank probe sensor connectors sheath filter vent valve toggle switch metal bracket tab