bd_facsaria_iii
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bd_facsaria_iii [2017/01/09 20:27] 85.246.171.69 |
bd_facsaria_iii [2018/06/08 18:01] flowcytometry |
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| <a href="/facility/flowcytometry/lib/exe/fetch.php?media=aria_iii_wiki.jpg"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=aria_iii_wiki.jpg" width=300></a> | <a href="/facility/flowcytometry/lib/exe/fetch.php?media=aria_iii_wiki.jpg"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=aria_iii_wiki.jpg" width=300></a> | ||
| <td style="border:0px solid white;"> <p style="line-height:1.8"> | <td style="border:0px solid white;"> <p style="line-height:1.8"> | ||
| - | <b>Location</b>: <a href="/facility/flowcytometry/lib/exe/fetch.php?media=flow_cytometry_map.jpg">Room P2-A-49</a> (<img src="/facility/bioimaging/lib/exe/fetch.php?media=telephone_icon.gif" width=15> 47222) <br> | + | <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>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-sorters/bd-facsaria-iii/m/744763/overview" target="_blank">FACSAria III</a><br> | <b>Model</b>: <a href="http://www.bdbiosciences.com/us/instruments/research/cell-sorters/bd-facsaria-iii/m/744763/overview" target="_blank">FACSAria III</a><br> | ||
| - | <b>Software</b>: FACSDiva<br> | + | <b>Nickname</b>: "Aria 3"<br> |
| - | <b>Year</b>: 2010 <br> | + | <b>Software</b>: FACSDiva 6.1.3<br> |
| + | <b>Year</b>: 2011 <br> | ||
| <b>SN</b>: P28200134 <br> | <b>SN</b>: P28200134 <br> | ||
|   <br> |   <br> | ||
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|   <br> |   <br> | ||
|   <br> |   <br> | ||
| - |   <br> | + | → <a href="https://imm.medicina.ulisboa.pt/intranet/booking/8"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=date.png"> BD FACSAria III Booking</a> |
| - | → <a href="https://imm.medicina.ulisboa.pt/intranet/booking/8"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png"> BD FACSAria III Booking</a> | + | |
| <br> | <br> | ||
| - | → <a href="/facility/flowcytometry/doku.php?id=bd_facsaria_iii_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png"> BD FACSAria III Usage Statistics</a> | + | → <a href="/facility/flowcytometry/doku.php?id=bd_facsaria_iii_usage"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=chart_line.png"> BD FACSAria III Usage Statistics</a> |
|   </p> |   </p> | ||
| </td> | </td> | ||
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| </table> | </table> | ||
| </html> | </html> | ||
| + | {{:info.gif|}} **Next Scheduled Maintenance**: October, 2018 | ||
| ===== System overview ===== | ===== System overview ===== | ||
| + | {{ :cell_sorting.png?0x300|}} The BD FACSAria III //cell sorter// is a high speed benchtop digital flow cytometer equipped with a sensitive, fixed-alignment cuvette flow cell. It has three spatially separated lasers - 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. Sorting can be performed into two or four 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 instrument is housed in a Baker BioProtect-IV bio-safety cabinet** and is able therefore to sort human samples. If you need a violet (405 nm) laser, check the [[bd_facsaria_iiu|BD FACSAria IIu]] instead. | ||
| + | |||
| + | * Three lasers with blue (488 nm), yellow-green (561 nm) and red (633 nm) | ||
| + | * Multicolor analysis of up to 10 parameters | ||
| + | * 70 μm, 85 μm and 100 μm nozzles available | ||
| + | * Up to 4-way sorting into 1.5 ml or 4 ml tubes and 2-way sorting into 15 ml tubes | ||
| + | |||
| + | ===== Configuration ===== | ||
| - | {{ :spinning_disk.jpg?0x300|}} The 3i Marianas SDC //spinning disk confocal microscope// is a fast imaging system which provides a trade-off between confocality, resolution and speed. It is an inverted microscope ideal for live cell applications which require fast acquisition speeds rather than high resolution images. The scanning unit achieves confocality by directing light through a spinning disk with many small pinholes. Images are then acquired with a sensitive EMCCD which allows for very small exposure times but is limited in resolution to 512x512 pixels. The stage is motorized and furthermore equipped with a piezo for Z displacement so fast 4D imaging is possible in multiple stage positions. The system is also equipped with a large size incubator for temperature control and a small stage incubator for CO2 supply. With this system you can perform optical sectioning of fluorescent samples which are too thick for a widefield system such as the [[zeiss_axio_observer|Zeiss Axio Observer]]. Even though its resolution is not as high as a //point-scanning confocal system// such as the [[zeiss_lsm_880|Zeiss LSM 880]] or [[zeiss_lsm_710|Zeiss LSM 710]], it is much faster than these two systems. If you need to use an upright microscope, check the [[zeiss_lsm_7_live|Zeiss LSM 7 Live]] fast confocal system instead. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]]. | + | {{:bd_facsaria_iii_configuration_wiki.png?800|}} |
| - | * **Microscope**: [[http://www.zeiss.com/microscopy/en_de/products/light-microscopes/axio-observer-for-biology.html|Zeiss Axio Observer]] | + | ===== Fluorophores Compatibility ===== |
| - | * **Confocal scanner**: [[http://www.andor.com/microscopy_systems/peripherals/confocal_scanners/|Yokogawa CSU-x1]] | + | |
| - | * **Camera**: [[http://www.photometrics.com/products/emccdcams/evolve/512.php|Evolve 512 EMCCD]] | + | |
| - | ===== System components ===== | + | {{:bd_facsaria_iii_fluorophores_wiki.png?700|}} |
| + | \\ | ||
| + | \\ | ||
| + | {{::bd_facsaria_iii_488.png?700|}} | ||
| + | \\ | ||
| + | {{::bd_facsaria_iii_561.png?700|}} | ||
| + | \\ | ||
| + | {{::bd_facsaria_iii_633.png?700|}} | ||
| - | ==== LASERs ==== | + | ===== Additional Information ===== |
| - | ^ Laser Unit ^ Wavelength ^ Maximum Power ^ Current Status ^ | + | |
| - | | Laserstack 488 | 488 nm | 100 mW | ok | | + | |
| - | | Laserstack 561 | 561 nm | 100 mW | ok | | + | |
| - | | Laserstack 640 | 640 nm | 100 mW | ok | | + | |
| - | ==== Objectives ==== | + | * **[[prepare_samples|How to prepare samples for Flow Cytometry]]** |
| - | ^ Magnification ^ Model ^ Type ^ NA ^ WD (mm) ^ | + | * **[[sorting_faq|What should I expect when sorting?]]** |
| - | | 20x | [[https://www.micro-shop.zeiss.com/?s=129353259d52f12&l=en&p=uk&f=o&a=v&m=s&id=420650-9901-000|Plan-Apochromat]] | Dry | 0.8 | 0.55 | | + | |
| - | | 63x | [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=420782-9900-000|Plan-Apochromat]] | Oil | 1.40 | 0.19 | | + | |
| - | | 100x | [[https://www.micro-shop.zeiss.com/?s=129353259d52f12&l=en&p=uk&f=o&a=v&m=s&id=420790-9901-000|Plan-Apochromat]] | Oil | 1.40 | 0.17 | | + | |
| - | **Upon request:** | + | ===== Booking Rules ===== |
| - | ^ Magnification ^ Model ^ Type ^ NA ^ WD (mm) ^ | + | |
| - | | 25x | [[https://www.micro-shop.zeiss.com/?s=129353259d52f12&l=en&p=uk&f=o&a=v&m=s&id=420852-9972-000|LCI Plan-NeoFluar]] | Oil/Glyc/W | 0.8 | 0.21 | | + | |
| - | | 40x | [[https://www.micro-shop.zeiss.com/?s=129353259d52f12&l=en&p=uk&f=o&a=v&m=s&id=420660-9970-000|Plan-Apochromat]] | Dry | 0.95 | 0.25 | | + | |
| - | ==== Emission Filtersets ==== | + | |
| - | ^ Setting ^ Transmission ^ | + | |
| - | | 1: 445 | 422-477 nm | | + | |
| - | | 2: 525 | 510-540 nm | | + | |
| - | | 3: 617 | 580-653 nm | | + | |
| - | | 4: 692 | 672-712 nm | | + | |
| - | | 5: QUAD | 440+521+607+700 nm | | + | |
| - | | 6: Block | No Transmission | | + | |
| - | | 7: Block | No Transmission | | + | |
| - | | 8: Block | No Transmission | | + | |
| - | | 9: Empty | Empty | | + | |
| - | | 10: 445 | 422-477 nm | | + | |
| - | ===== Microscope Turn On Procedure ===== | + | The system is available for booking from 10am to 10pm from Monday to Friday. |
| - | * Check that the main power supply switch is on (should be on by default) | + | |
| - | {{marianas_power1.jpg?|}} | + | |
| - | * Turn on the fluorescence lamp (if needed) | + | |
| - | {{marianas_power5.jpg?|}} | + | |
| - | * Turn on the secondary power supply switch | + | |
| - | {{marianas_power2.jpg?|}} | + | |
| - | * Turn on the power switch distributor (green) and the component power switches (red) **one at a time** | + | |
| - | {{marianas_power3.jpg?|}} | + | |
| - | * Turn on the Yokogawa spinning disk scanner (using the key) | + | |
| - | {{marianas_power4.jpg?|}} | + | |
| - | * Turn on incubation and CO2 on the touchscreen (if needed) | + | |
| - | {{marianas_power6.jpg?|}} | + | |
| - | * Turn on the lasers you need to use (using the key and the power switches) | + | |
| - | {{marianas_power7.jpg?|}} | + | |
| - | * Turn on the computer | + | |
| - | * Wait 2 min to allow for system components detection | + | |
| - | * Start the **SlideBook** software | + | |
| - | **Warning**: If you need to change the stage adaptor, please contact the <html><a href="/facility/bioimaging/doku.php?id=about">Bioimaging Facility</a> (<b>imm-bioimaging@medicina.ulisboa.pt</b> | <img src="/facility/bioimaging/lib/exe/fetch.php?media=telephone_icon.gif" width=15> 47316</html>) | + | 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. | ||
| - | ===== Microscope Turn Off Procedure ===== | + | Please check the {{ ::sop.ucf.004_-_ucf_imm_user_guidelines.pdf |User Guidelines}} for detailed information. |
| - | * Remove your sample from the stage/piezo holder | + | |
| - | * Exit the **SlideBook** software | + | |
| - | * Switch off the computer | + | |
| - | * Turn off all system components (follow the Turn On Procedure in reverse) | + | |
| ---- | ---- | ||
| [[resources|Back to the Resources page]] | [[resources|Back to the Resources page]] | ||
bd_facsaria_iii.txt · Last modified: 2024/06/10 23:03 by flowcytometry
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