Flow Cytometry Wiki

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<b>Location</b>: <a href="/facility/flowcytometry/lib/exe/fetch.php?media=flow_cytometry_map.jpg">Room P2-A-49</a> (<img src="/facility/flowcytometry/lib/exe/fetch.php?media=telephone_icon.gif" width=15> 47224) <br> 
<b>Manufacturer</b>: <a href="http://www.bdbiosciences.com/eu/home" target="_blank">BD Biosciences</a><br>
<b>Model</b>: <a href="https://www.bdbiosciences.com/en-us/products/instruments/flow-cytometers/research-cell-analyzers/bd-facsymphony-a5-se" target="_blank">BD FACSymphony A5 SE</a><br>
<b>Nickname</b>: "A5 SE"<br>
<b>Software</b>: FACSDiva 9.0<br>
<b>Year</b>: 2023 <br>
<b>SN</b>:  <br>
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&rarr; <a href="https://my.agendoscience.com/calendar/?id=MTk2Y2FsZW5kYXJz" target="_blank"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=booking.png">  BD FACSymphony A5 SE Booking</a>
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&rarr; <a href="/facility/flowcytometry/doku.php?id=bd_facsymphony_usage"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=statistics.png"> BD FACSymphony Usage Statistics</a> 
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&rarr; <a href="https://imm.medicina.ulisboa.pt/facility/flowcytometry/lib/exe/fetch.php?media=bd-facsymphony-a5-se-brochure1.pdf" target="_blank"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=brochure.png">  BD FACSymphony A5 SE Brochure</a>
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{{:info.gif|}} **Next Scheduled Maintenances**:

{{:info.gif|}} [[aria_iii_contamination|Pseudomonas contamination timeline]]

===== System overview =====

{{  :cell_sorting.png?0x300|}} The BD FACSymphony S6 SE cell sorter is a high speed benchtop digital flow cytometer equipped with a sensitive, fixed-alignment cuvette flow cell. It has five spatially separated lasers - 349 nm, 405 nm, 488 nm, 561 nm and 633 nm. Its primary function is to analyze complex populations of cells and yield pure populations of cells meeting defined criteria. The sorter can purify samples that are simply positive and negative for a single fluorophore or as complex as multi-color samples with complex gating strategies.

The BD FACSymphony™ A5 SE allows for high-parameter spectral flow cytometry. It enables you to gain full visible spectrum coverage to collect all light emitted without the need for filter changes, increasing dye flexibility and number of simultaneous colors. It also allows for autofluorescence unmixing, that may enable the resolution of difficult cells and populations.


Sorting can be performed into four or six tubes with speeds up to 25,000 events/second. The system sorts by incorporating cells from the sample tube into a stream of sterile PBS. The stream is interrogated by the lasers at the flow cell and the system electronics keeps track of each cell as they pass through the laser and determines specific cells that meet the sort criteria. A transducer vibrates the stream and induces droplet formation, with cells in the stream being incorporated into the droplets. If a cell meets the sorting criteria and is in the last drop before the break off, the instrument will charge that drop. The charged droplet is then deflected into the proper collection tube by the charge plates. Different cell types can be sorted with the use of 70 μm, 85 μm and 100 μm nozzles. **The fully integrated Class II Type A2 biosafety cabinet meets personal and product protection, reducing the risk of operator exposure, while sorting potentially hazardous samples (e.g human samples and other BSL-2 level samples).**
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  * 48 fluorescent detectors
  * Five lasers with ultra-violet (349nm), violet (405 nm), blue (488 nm), yellow-green (561 nm) and red (640 nm) 
  * Enables both spectral unmixing and compensation workflows
  * 70 μm, 85 μm and 100 μm nozzles available
  * Up to 6-way sorting into 1.5 ml or 4 ml tubes and 4-way sorting into a combination of 4 mL tubes and 15 ml tubes
  * Sorting for 6-, 12-, 24-, 96-, 384-well, PCR and other custom plates
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===== Configuration =====
{{:facsymphony_optical_configuration.png?900|}}
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===== Suggested Fluorochrome Peak Channels =====

{{{{:facsymphony_fluorophores.png?900|}}
{{{{:facsymphony_fluorophores_2.png?900|}}

===== Useful Links when Preparing your Experiment =====

  * [[prepare_samples|{{how_to_prepare_samples_for_flow_cytometry.png}}]]
  * <html><a href="http://www.bdbiosciences.com/en-us/applications/research-applications/multicolor-flow-cytometry/product-selection-tools/spectrum-viewer" target="_blank"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=bd_spectrum_viewer.png" WIDTH=300></a></html>
  * <html><a href="http://www.bdbiosciences.com/en-us/applications/research-applications/multicolor-flow-cytometry/product-selection-tools/spectrum-viewer" target="_blank"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=bd_research_cloud.png" WIDTH=300></a></html>

===== Additional Information =====

  * [[https://imm.medicina.ulisboa.pt/facility/flowcytometry/lib/exe/fetch.php?media=sop.ucf.038_-_bd_facsymphony_a5_se_operation.pdf| BD FACSymphony A5 SE Operation]]
  * [[https://imm.medicina.ulisboa.pt/facility/flowcytometry/lib/exe/fetch.php?media=sop.ucf.022_-_bd_lsrfortessa_x-20_operation.pdf| BD FACSDiva Tutorial (for BD FACSymphony A5 SE)]]	 
  * [[https://imm.medicina.ulisboa.pt/facility/flowcytometry/lib/exe/fetch.php?media=sop.ucf.024_-_bd_lsrfortessa_facsymphony_a5_se_troubleshooting.pdf| BD LSRFortessa & FACSymphony A5 SE Troubleshooting]]
  * [[https://imm.medicina.ulisboa.pt/facility/flowcytometry/lib/exe/fetch.php?media=sop.ucf.040_-_bd_facsymphony_cleaning.pdf| BD FACSymphony A5 SE Cleaning]]
  * [[https://imm.medicina.ulisboa.pt/facility/flowcytometry/lib/exe/fetch.php?media=sop.ucf.036_-_bd_facsymphony_a5_se_starting_up_cleaning.pdf| BD FACSymphony A5 SE Starting up Cleaning]] for the first user of the day, on weekends and out of staff working hours 

===== Booking Rules =====

The system is available for booking from 10am to 6pm on Monday, Wednesday and Friday. 

The cell sorting service is provided by a dedicated technician upon reservation on the online booking system. 
The system can be booked up to 2 months in advance. 

Please check the {{ ::sop.ucf.004_-_ucf_imm_user_guidelines.pdf |User Guidelines}} for detailed information.

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