Team:SUSTech Shenzhen/InterLab

Interlab

Contribution

Flow Cytometer Measurement

Materials

• 194.7 g FITC (provided in kit)
• 10ml 1xPBS (phosphate buffered saline) 96 well plate
• Competent cells (Escherichia coli strain DH5α)
• LB (Luria Bertani) media with Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH), 1 ml Falcon tube for cell growth Incubator at 37°C, 1.5 ml eppendorf tubes for sample storage Ice bucket with ice,Pipettes, SpheroTech Rainbow Calibration Particles RCP-30-5A, CytoFlex flow cytometer

Devices (from InterLab Measurement Kit):

• Positive control
• Negative control
• Device 1: J23101+I13504
• Device 2: J23106+I13504
• Device 3: J23117+I13504

Methods

1. Open computer, click cytometer setting, load clean solution and system startup program for initialization.

2. Load QC(Lot: 45065) falcon tube to do pre-tests.

3. Load Rainbow beads, set FSC-A-SSA, FSC-A-FSC-H, FITC-A-Count.

4. Mix 100ul overnight culture with PBS, load samples and examine the fluorescence.

5. Close experiment and perform daily clean with ddH2O. Exit the software.

Result

Tab. 1 Result of flow cytometer measurement

Fig. 1 FITC fluorescence peaks of Rainbow beads

Fig. 2 FITC fluorescence peak of negative control

Fig. 3 FITC fluorescence peak of positive control

Fig. 4 FITC fluorescence peak of Device 2

Made by from the iGEM team SUSTech_Shenzhen.

Licensed under CC BY 4.0.

• ©2016 SUSTech_Shenzhen