Difference between revisions of "Team:SUSTech Shenzhen/InterLab"

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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)
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* 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
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== Methods ==
 +
<ol style="list-style-type: lower-alpha;">
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<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>
  
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== Result ==
 +
 +
[[File:media/image1.png|649x432px]]
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 +
Tab. 1 Result of flow cytometer measurement
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 +
[[File:media/image2.png|368x381px]]
 +
 +
Fig. 1 FITC fluorescence peaks of Rainbow beads
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 +
[[File:media/image3.png|368x381px]]
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Fig. 2 FITC fluorescence peak of negative control
 +
 +
[[File:media/image4.png|368x382px]]
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Fig. 3 FITC fluorescence peak of positive control
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[[File:media/image5.png|368x382px]]
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Fig. 4 FITC fluorescence peak of Device 2
  
 
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Revision as of 05:06, 17 October 2016

Team SUSTC-Shenzhen

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

649x432px

Tab. 1 Result of flow cytometer measurement

368x381px

Fig. 1 FITC fluorescence peaks of Rainbow beads

368x381px

Fig. 2 FITC fluorescence peak of negative control

368x382px

Fig. 3 FITC fluorescence peak of positive control

368x382px

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