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1 2008/02/05 Isoiso
2 FACSCalbur 488nm FL1 530/30 FITC GFP alexa488 FL2 585/42 PE Cy3 PI FL3 650LP Percp PC5 PI PE-C 7 Red APC Falcon BD Calibrite 3 BD No DNA QC particles BDNo Flow-Cheak(10um) BeckmanColter Flow-Set(3.6um) BeckmanColter HI SPHEROTM UltraRainbow Fluorescent Particles A single population of Rainbow Fluorescent Particles with enhanced UV and Far Red fluorescence intensity. URFP-30-2 ; UltraRainbow Fluorescent Particles, 10 7 /ml, 2 ml, m, $ URFP-38-2 ; UltraRainbow Fluorescent Particles, 10 7 /ml, 2 ml, m, $ URFP ; UltraRainbow Fluorescent Particles, 10 7 /ml, 2 ml, m, $ URFP ; UltraRainbow Fluorescent Particles, 1% w/v, 5 ml, m, $ Spherotech Rainbow Beads-8peek
3 FACSCallibur FACSCallibur ON ON ON FACSCallibur PRIME STANDBYPRIME STANDBY FACSComp FACSCallibur (SIP) (DCM) Run (SIT) HI FACScanSTANDBY standby10facscaliburoff
4 MAX 3/4 2 1 OFF PUSH Line
5 CRT G4 1.ON 2.caliburON 10 CRT G4 3.ON 4.MAC ON 1
6 1. 2.ON Run Push 3. 4.LINE close OPEN
7 5.TUBE LOW MED HI RUN STANDBY PRIME 6-1.Prime 6-2.StandbyPrime 6-3.StandbyPrime 7.Standby1TUB 3 30 Warm 10OFF 8.CellQuest STATAS Standby.MacellQuest USBSOFTUSB +B calibur Detecters/Amps Threshold Compensation Stats
8 CellQuest USBSOFTUSB MacellQuest 1.dataFolder +B calibur Detecters/Amps Threshold Compensation Stats Folder 1-3.DeskTop USB MO COPY COPY USB WindowsMO data WIN-MAC File MAC SOFT FACS File 1-4.New 1-6.Select 1-7.Parameter Description 2.Counter 1-5. Create
9 2 FITC PE IsoType DotPlot Acquisit**->Analysis FSC-H SSC-H 55 Ok DotPlot FL1-H 55 FL2-H Ok Acquisit**->Analysis 55 FL1-H Ok FL2H 55 Ok
10 1.STANDBY 2.Detecters/Amps LOW MED H I RUN STANDBY PRIME 3.Run Low HI 4.Acquire Pause Abort A uaire data 5.FSC/SSC dot ( 6. (Cal bure0427 Mode Lin Voltege ModeLog PE 7.FL1/FL FITC 10 1 FL1/FL2dotPlot Threshold Voltege
11 FITC 1. FITC LOW MED H I RUN STANDBY PRIME 2.Run Low HI 3.Acquire Pause Abort A uaire data FL2-FL1 FL2-FL1 Good PE 1. PE LOW MED H I RUN STANDBY PRIME 2.Run Low HI 3.Acquire Pause Abort A uaire data FL1-FL2 FL1-FL2 Good
12 OKdata OK 4. LOW MED H I data RUN STANDBY PRIME 5.Run LowHI 6. 7.Acquiredata data
13 FCS2.0data Plotdata Cell Quest File----New Gate List GateRotate Region command Overlay/stats/
14 Overlay/stats/ ( 1. ControlGate Gate GatePlotFormat R1
15 Analysis Selectdata Overlay-- Histogram K-S Region Gate plot 4
16 DNA CellCyclePlot PI AADFL2 FL3 3 1.Lin 2.FL2 3.Plot DotPlot Acquisit**->Analysis FSC-H SSC-H Ok DotPlot FL2-W FL2-A Ok Acquisit**->Analysis SSC-H Ok FL2A Ok
17 Cell ycle 1. PI LOW MED H I RUN STANDBY PRIME 2.Run Low LOW MAX400cells/sec Pause Abort A uaire data 3.Acquire Cell SSC200 FL2 FL2-A G0/G1 200 A-W single
18 ThresholdFL2 24 OKdata OK 4. LOW MED H I data RUN STANDBY PRIME 5.Run LowHI 6. 7.Acquiredata data
19 FL2A/W
20 BD No BD Calibrite 3 11 L 1 13 L OK TAgetValue UN MIX soft 1.FACSComp 2.Run 4.LotID ID Fail 3.Lyse/Wash 5.Run 6.UN UN 7.Run HI HI LOW MED H I RUN STANDBY PRIME 8.Start
21 Rate Run HI HI LOW MED H I RUN STANDBY PRIME 11.Start PASS PASS ID Line 3-Color Lyse/No-Wash FACSComp Report 12.Quit 2 Institution: Date: :29 PM Director: Software: FACSComp 4.2 Operator: iso Cytometer: FACSCalibur E0427 Parameter High Low Separation Minimum Result Lot ID FSC Pass 31845I SSC Pass 31845I FL Pass 27903O FL Pass 29497J FL Pass 34254M Parameter Detector Amplifier Threshold Blue Laser Current 5.94 Amps FSC E Blue Laser Power mwatts SSC FL1 554 Log FL2 468 Log FL3 571 Log 300 Compensation FL1-%FL2 FL2-%FL FL2-%FL3 0.0 FL3-%FL URL Comments:
22 Shutdown 1.Prime LOW MED HI RUN STANDBY PRIME ( 2.5cm LOW MED HI RUN STANDBY PRIME RUN--HI RUN 3.Prime LOW MED HI 3-1 RUN STANDBY PRIME Standby
23 cm LOW MED HI RUN STANDBY PRIME RUN--HI RUN 5. 1( 1cm) 5-1 OFF 1 6.STANDBY LOW MED HI RUN STANDBY PRIME. 8.Stand By10
24 1 Pt Plot Plot Fig HD Fil PICT.
25 File File instrument data Done
26
27
28 LOW MED H I RUN STANDBY PRIME run Statas 15 WNot Ready Sheath Fiuid:Empty Waste Tank:full
29 Falcon um 100um1/3 A 1cmSIP LOW MED H I RUN STANDBY PRIME Prime FACS Drin--Fill backfrash 1010RUNHI B C 10 68
30
31 CELLQuest : Compensation
32
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