BD Biosciences FACSMelody
Quick Facts
- Biosafety level 2 cabinet
- 3 lasers, 11 parameters*
405, 488, 640 nm lasers
- 100 um nozzle
- Controlled sample input/output temperature
- High-speed 2-way sorting
- High-throughput plate sorting
* includes FSC and SSC in addition to colors
Note: Users must use Falcon tubes (Cat# 352008 or 352235 with filter cap) or Flow Cytometry tubes from MTC Bio (https://mtcbiotech.com/product/flowtubes-for-flow-cytometry-instruments/). MTC Bio strainer tubes are not as good as Falcon.
Sorting Guideline – BRC Flow Facility
Location: Biotech 140
Technical Specifications
The BD FACSMelody is easy to use as its streamlined workflow eliminates manual setup of the stream and monitoring of the sort, which minimizes hands-on time. Unlike the FACSAria Fusion, learning to operate the Melody is easy. The BD FACSChorus software prompts novice users with step-by-step instructions and it eliminates the need for setting up the sort stream and monitoring the sort, reducing the risk of poor sort outcome. The Melody has a fixed flow cell and reusable nozzle. Sort devices include 2 -way tubes as well as 6-, 24-, 48-, 96-, and 384-well plates. Additional accessories include biological safety cabinet and aerosol management system.
| Excitation lasers | 488 nm, 640 nm, 405nm |
| Output power | 405 nm (36 mW), 488 nm (16 mW), 640 nm (36 mW) |
| Detection parameters | 9 fluorescence + 2 scatter |
| Sample tube | Single, auto-loading tube |
| Tube types | 5.0mL |
| Temperature control | 5°C, 37°C |
| Agitation unit | Eccentric rotation |
| Magnetic drive | 300 rpm speed |
| Sorting nozzle | 100 um |
Instrument Configuration
| Laser | Filter BP | Filter LP | Preferred colors |
|---|---|---|---|
| 405 | 448/45 | Pacific Blue, V450, DAPI, Brilliant Violet 421 | |
| 528/45 | 500 | AmCyan, V500, Brilliant Violet 510 | |
| 738/36 | 752 | Brilliant Violet 786 | |
| 488 | 527/32 | 507 | Alexa Fluor 488, BB515, GFP, FITC |
| 586/42 | 569 | PI, PE | |
| 700/54 | 665 | 7-AAD, PE-Cy5, PerCP-Cy5.5, PerCP | |
| 783/56 | 752 | PE-Cy7 | |
| 640 | 660/10 | Alexa Fluor 647, APC | |
| 783/56 | 752 | APC-H7, APC-Cy7 |
Saving Your Data
Users are given access to a fileshare drive to save data after being trained on the instrument. You can save your data as FCS files, sort reports, dot plots, histograms, etc.
FCS Files:
1. Under the View Data tab, under Data Sources -> Export FCS files.
2. Save to This PC -> Cytometry network drive, under your folder. Will save as .zip folder.
Sort Reports:
1. Sort reports are only generated for experiments that have done sorting.
2. Under View Reports tab -> Select the sort from the dropdown list -> Export Report.
3. Save to This PC -> Cytometry network drive. Will save as .pdf
4. Repeat for each sort file on the dropdown.
To save Dot plots, Histograms, etc:
1. Take screenshots by pressing Ctrl + P. This will save a pdf of everything on the screen.
2. Save to This PC -> Cytometry network drive. Repeat for any files you wish to have screenshots.