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="http://www.bdbiosciences.com/us/instruments/research/cell-analyzers/bd-lsrfortessa/m/751752/overview" target="_blank">LSRFortessa</a><br>
<b>Nickname</b>: "Fortessa 2"<br>
<b>Software</b>: FACSDiva 6.2<br>
<b>Year</b>: 2010 <br>
<b>SN</b>: H5B300009 <br>
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&rarr; <a href="https://imm.medicina.ulisboa.pt/intranet/booking/73"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=date.png">  BD LSRFortessa 2 Booking</a>
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&rarr; <a href="/facility/flowcytometry/doku.php?id=bd_lsrfortessa_2_usage"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=chart_line.png">  BD LSRFortessa 2 Usage Statistics</a>
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{{:info.gif|}} **Next Scheduled Maintenance**: December, 2018

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

{{  :cell_analyzers.png?0x300|}} The BD LSRFortessa 2 //cell analyzer// is a benchtop digital flow cytometer equipped with a fixed-alignment flow cell that is gel-coupled to the collection optics. It has three spatially separated lasers - 405 nm, 488 nm and 640 nm. Its primary function is to characterize complex populations of cells. Individual cells pass single file through a stream, which is interrogated by the three lasers with the emitted light being detected by photomultiplier (PMT) tubes. The system electronics calculates the time it takes a single cell to pass through each laser intercept and assigns the signal of each PMT to the appropriate cell. Besides fluorescence, the physical properties of the cell are also analyzed with side scatter (SSC), which reflects the internal complexity of a cell and forward scatter (FSC), which is roughly proportional to cell size. The sample flow rate can be set to low, medium and high values, which correspond approximately to 12, 35 and 60 μl/minute, respectively. The LSRFortessa 2 is equipped with a **High Throughput Sampler (HTS)**, which allows the automated acquisition of samples from 96 and 384 well plates. If your cells are stained with PerCP, check the [[bd_lsr_fortessa_1|BD LSRFortessa 1]] instead. If you need multicolor analysis of up to 16 parameters, check the [[bd_lsr_fortessa_x20|BD LSRFortessa X-20]].

  * Three lasers with violet (405 nm), blue (488 nm) and red (640 nm)
  * Multicolor analysis of up to 10 parameters

===== Configuration =====

{{:bd_lsrfortessa_2_configuration_wiki.png?800|}}

===== Fluorophores Compatibility =====

{{:bd_lsrfortessa_2_fluorophores_wiki.png?700|}}
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{{::bd_lsrfortessa_additional_filters.png?220|}}
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{{::bd_lsrfortessa_2_405.png?700|}}
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{{::bd_lsrfortessa_2_488.png?700|}}
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{{::bd_lsrfortessa_2_640.png?700|}}

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

  * **[[prepare_samples|How to prepare samples for Flow Cytometry]]**
  * [[https://imm.medicina.ulisboa.pt/files/5815/4825/3980/FORM.UCF.043_-_BD_LSRFortessa_1_and_2_operation.pdf| BD LSRFortessa Operation]]
  * [[https://imm.medicina.ulisboa.pt/files/5515/4827/1692/FORM.UCF.047_-_BD_LSRFortessa_troubleshooting.pdf| BD LSRFortessa Troubleshooting]]
  * [[https://imm.medicina.ulisboa.pt/files/5114/8700/1772/FORM.UCF.052_-_HTS_Operation_for_users.pdf| High Throughput Sampler (HTS) Operation]]

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

Available 7 days a week, 24 hours a day for trained researchers. 

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

===== Cleaning Protocol =====

{{::cleaning_procedure_fortessa.png?700|}}

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