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| + | = 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 == | ||
| + | <ol style="list-style-type: lower-alpha;"> | ||
| + | <li><p>Open computer, click cytometer setting, load clean solution and system startup program for initialization.</p></li> | ||
| + | <li><p>Load QC(Lot: 45065) falcon tube to do pre-tests.</p></li> | ||
| + | <li><p>Load Rainbow beads, set FSC-A-SSA, FSC-A-FSC-H, FITC-A-Count.</p></li> | ||
| + | <li><p>Mix 100ul overnight culture with PBS, load samples and examine the fluorescence.</p></li> | ||
| + | <li><p>Close experiment and perform daily clean with ddH2O. Exit the software.</p></li></ol> | ||
| + | == Result == | ||
| + | {{SUSTech_Image_Center_12 | filename=T--SUSTech_Shenzhen--B74F8563-C2AD-497D-97E7-5F96FB3B3034.png | caption=Tab. 1 Result of flow cytometer measurement | width=2000px}} | ||
| + | {{SUSTech_Image_Center | filename=T--SUSTech_Shenzhen--851A11BD-5BF8-4D19-A68D-2DBAF254C59F.png | caption=Fig. 1 FITC fluorescence peaks of Rainbow beads | width=1000px}} | ||
| + | {{SUSTech_Image_Center | filename=T--SUSTech_Shenzhen--73D6A3C0-5B40-4E21-B604-8D6A4B81F698.png | caption=Fig. 2 FITC fluorescence peak of negative control | width=1000px}} | ||
| + | {{SUSTech_Image_Center | filename=T--SUSTech_Shenzhen--9247196A-FE86-4BF7-98C5-9BEF806BAEB1.png | caption=Fig. 3 FITC fluorescence peak of positive control | width=1000px}} | ||
| + | {{SUSTech_Image_Center | filename=T--SUSTech_Shenzhen--FAA88036-4284-4532-9C21-2479A04DE1A8.png | caption=Fig. 4 FITC fluorescence peak of Device 2 | width=1000px}} | ||
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Latest revision as of 23:13, 19 October 2016
Interlab
Contribution
Contents
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
Open computer, click cytometer setting, load clean solution and system startup program for initialization.
Load QC(Lot: 45065) falcon tube to do pre-tests.
Load Rainbow beads, set FSC-A-SSA, FSC-A-FSC-H, FITC-A-Count.
Mix 100ul overnight culture with PBS, load samples and examine the fluorescence.
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
