Session times: 9:30 - 5:00, or by appointment. Allow for 30min machine setup + 25 min per sample when booking. |
Software dongles are available for loan. The dongles can be found on level 9.
Please do not hold these dongles overnight unless confirmed with a Detmold Staff member that the dongle is not required for installation or training in the morning. Please do not take these dongles off site, i.e. they are never to be taken home for use. If the dongle is lost the cost of replacing the software licence will be billed according to its loss.
A fair use policy applies to all of the software dongles. Please be aware that Detmold Staff may contact you for return of the dongle if your usage has exceeded a fair limit.
FCS express v7 New | Internet licence. Windows 7/8/10/11; macOS 10.12+. Can be used for offline analysis of flow data from any analyser or sorter. Cytoflex data needs manual adjustment (see cytoflex on the instruments page). Note: does not have cell cycle or proliferation modules. |
FCS express v6 | Older version of FCS express. Works on windows 7/8/10/11; macOS upto 10.14. Can analyse data from any sorter or analyser. Cytoflex data needs manual adjustment (see cytoflex on the instruments page). This licence includes the modules for cell cycle analysis and proliferation analysis. |
FCS express v4 | Older version of FCS express (replaced by v6 in Jan 2017). Does not have access to additional modules. |
FCAP Array | Software for analysis of Cytometric Bead Arrays (CBA) from BD |
Kaluza | Software package released by Beckman Coulter for offline analysis |
FlowJo | Licences can be purchased from SAHMRI. |
User category | CCB Staff | External academic | Commercial |
---|---|---|---|
Cell sorting | $60/hr | $100/hr | $200/hr |
Northern Lights | $42.5/hr | $42.5/hr | $120/hr |
Fortessa | $30/hr | $50/hr | $120/hr |
Gallios / Cytoflex | $20/hr | $35/hr | $70/hr |
Operator-fee | $30/hr | $40/hr | in-price |
Sort tubes | $7.95 per bag of 25 | ||
Analysis tubes | $15.05 per bag of 500 | ||
Analysis-tube lids | $20.75 (1000pc) $8.67 (250pc) | ||
non-sterile filter BD 30 µm | $2.6 each (with 1mL syringe) | ||
Confocals | $28/hr | $60/hr | $240/hr |
STED | $28/hr | $50/hr | $240/hr |
The Gallios has 4 lasers (405nm, 488nm, 561nm, 640nm) and is capable of analysing 10 fluorochromes.
It is carousel-based, with each carousel holding 32 tubes.
CCB Gallios guide.pdf | Download (1.4 Mb) |
Channel | Optical filter | Fluorochromes (405nm/488nm/640nm) | with 561nm laser ON |
Pick only one fluorochrome from each row. *the 488 and 561 lasers share FL1-FL5 detetors. | |||
FL1 | 525 BP 40 | FITC, GFP, CFSE, Alexa 488, Calcein, DyLight 488, Live/Dead Green | - |
FL2 | 575 BP 30 | PE | Alexa 546, eFluor 585 NC, dsRed, tdTomato |
FL3 | 620 BP 30 | PE-Texas Red, ECD, PI | mCherry, Alexa 568 |
FL4 | 695 BP 30* | 7-AAD, PerCP, PerCP-Cy5.5, PE-Cy5, PE-Cy5.5, PerCP-eFluor710 | (675/30): mPlum |
FL5 | 775 LP | PE-Cy7 | - |
FL6 | 660 BP 20 | APC, Alexa 633, Alexa 647, eFluor 660, Live/Dead Far Red | |
FL7 | 725 BP 20 | APC-Alexa 700 | |
FL8 | 755 LP | APC-Cy7, APC-H7, APC-Alexa750, APC-eFluor780 | |
FL9 | 450 BP 50 | Pacific Blue, Alexa 405, Creulean, BV421, Horizon V450, DAPI, Hoechst, eFluor 450 | |
FL10 | 550 BP 40 | Krome Orange, FluoroGold, Pacific Orange, Alexa 430, BV 570, Cascade Yellow, Live/Dead Aqua | |
* 695 BP 30 is the standard filter and can be swapped for 675 BP 30 to maximise signal from certain fluorochromes | |||
This list is an example of commonly used fluorochromes. For reagents not on the list, facility staff can help determine compatability. |
The Fortessa has 5 lasers and can simultaneously analyse 18 fluorochromes
355nm (20 mw), 405nm (50 mw), 488nm (50 mw), 561nm (50 mw), 633nm (40 mw)
Fortessa fluorochrome options.pdf | Download |
Fortessa standard configuration.pdf (updated April 2020) |
Download |
CCB Fortessa guide.pdf | PDF | PDF(dark) |
Fortessa panel builder (std config).xlsx | XLSX file |
Laser | Filter |
---|---|
355 nm 20 mW |
379/28 |
515/30 | |
740/35 | |
405 nm 50 mW |
450/50 |
525/50 | |
610/20 | |
660/20 | |
710/50 | |
780/60 | |
488 nm 50 mW |
488/10 (SSC) |
530/30 | |
710/50 | |
561 nm 50 mW |
585/15 |
610/20 | |
670/30 | |
780/60 | |
640 nm 40 mW |
670/14 |
730/45 | |
780/60 |
This badboy has 5 lasers (355nm, 405nm, 488nm, 561nm, 640nm). It can analyse 17 fluorochromes.
Sorting formats: 6-way 5 mL-tube, plate-sorting and small particle sorting.
Astrios panel builder | astrios panel builder.xlsx |
Astrios complete detector layout | Detector setup.pdf |
Spare filters | 448/59, 513/26, 546/20, 610/20, 614/20, 620/29, 664/22, 670/30 | 600 LP, 650 LP |
Laser | Filter | Common flurophores |
---|---|---|
355 nm 80 mW |
422/44 | BUV395; DAPI (448/59) |
505/40 | BUV496; BUV563 (546/22) | |
722/44 | BUV737; BUV661 (664/22) | |
405 nm 20 mW |
405/10 | SSC (small particles) |
448/50 | Cerulean; BV421; Pacific Blue | |
526/52 | BV500; mT-Sapphire | |
630/40 | BV605; BV650 (664/22); FVS-575v | |
785/50 | BV786 | |
488 nm 100 mW |
488/6 | SSC |
513/26 | GFP; FITC; CFSE; AF488 | |
576/21 | ||
692/75 | PE-Cy5.5; PerCP; PerCP-eFluor 710 | |
797/70 | PE-Cy7 | |
561 nm 110 mW |
561/4 | SSC |
579/16 | DsRed; tdTomato; PE | |
620/29 | NY610; mCherry; PI | |
692/75 | PE-cy5; NY660; mPlum; 7-AAD | |
640 nm 60 mW |
642/10 | SSC |
671/30 | APC; AF647; SytoxRed | |
710/45 | APC-R700; Zombie-NIR | |
797/70 | Cy7; APC-Cy7; APC-eFluor780 |
This machine is no longer onsite, so arrangements should be made to run samples on the Astrios instead.
The Aria has 3 lasers (407nm, 488nm, 633nm) and is capable of analysing 8 fluorochromes simultaneously.
Sorting formats: 4-way 5 mL-tube and plate-sorting.
The lack of a 561-laser means that experiments involving RFP/mCherry should be run on the Astrios instead.
Laser | Mirror | Filter | Common flurophores |
---|---|---|---|
407 nm | - | 450/40 | Alexa Fluor 405, BV421, Cascade Blue, CFP, DAPI, Hoechst, Hoechst Blue, Indo-1 (Blue or Violet), Pacific Blue, V450, BFP, Cerulean, eCFP, Hoescht 33342, Live/Dead Violet |
502 | 530/30 | Alexa Fluor 430, BV510, Pacific Orange, V500, Krome Orange, Live/Dead Aqua, SYTOX Blue | |
690 | 710/50 | BV650, BV711, BV786, Hoechst Red, Qdot 525, Qdot 705, Qdot 800, FG, FluoroGold, UVR | |
488 nm | - | 488/10 | SSC |
502 | 530/30 | Alexa Fluor 488, FITC, GFP, Calcein, CFSE, Cy2, DyLight 488, Emerald, Live/Dead Green, SYBR Green, YFP | |
556 | 585/42 | PE, Alexa Fluor 546, Alexa Fluor 555 | |
610 | 616/23 | PE-Texas Red, Alexa Fluor 568, BD Horizon PE-CF594, Belgian Red, ECD, Propidium Iodide | |
655 | 695/40 | 7-AAD, PE-Cy5, PE-Cy5.5, PerCP, PerCP-Cy5-5, PI, PerCP-eFluor 710, PerCP-Vio770 | |
735 | 780/60 | PE-Cy7 | |
633 nm | - | 660/20 | Alexa Fluor 680/700/750, APC-Cy7, APC-H7, APC-AlexaFluor 700/750/780, APC-Vio770 |
735 | 780/60 | APC, Alexa Fluor 647, Alexa 647, Alexa Fluor 633, Cy5, Cy5-5, DRAQ5, Live/Dead Far Red |
Manufacturer links have been depreciated. However, there are resources available online via your favourite search engine (search for: FACSAria II).
The LSM 700 confocal microscope is located in the Detmold facility. For access and training please contact us.
The LSM 710 two-photon microscope is located in the basement. Please contact Claudine Bonder for access and training.
The Leica SP8 is located on the third floor. Please contact Michael Samuel for access and training.
The histech confocal slide scanner is located in the genomics facility (0-46-N). Please contact Michael Samuel for access and training.
The LSM 800 confocal microscope is located in the Detmold facility. For access and training please contact us.
The Cytek Northern LightsTM-Clinical (NL-CLC) 3000 is a full spectrum flow cytometry system capable of up to 41 detection channels (38 fluorescence channels, FSC, blue laser SSC and violet laser SSC). The system is suitable for dyes emitting in the 420 – 829 nm range.
It has three lasers 405 nm (100 mW), 488 nm (50 mW), 640 nm (80 mW). The detector modules include violet: 16 channels from 420 nm – 829 nm, blue: 14 channels from 498 – 829 nm, and red: 8 channels from 652 – 829 nm.
Machine minder | Sheree Bailey, Bradley Chereda |
Manufactuer product page | Link |
User guide | Link (PDF) |
Fluorochrome selection guide | Link (PDF) |
Full spectral viewer | Link |
The Cytoflex has 2 lasers (488nm and 561nm) and is capable of analysing 6 fluorochromes.
Tube format: 5mL (12x75) / 1.5mL(microcentrifuge).
The cytoflex is pump-based/volumetric and records fluorescence data at a 7-log resolution.
Cytoflex manufactuers link | Link |
Cytoflex manual | Link |
Cytoflex config | PDF (286 kb) |
Cytoflex FCS express 6 template | .fey template file (62 kb) |
Cytoflex FCS express 7 template | .fey template file (28 kb) |
Optical filter | Fluorochromes |
525 BP 40 | FITC, GFP, CFSE, Alexa 488, Calcein, DyLight 488, Live/Dead Green |
690 BP 50 | 7-AAD, PerCP, PerCP-Cy5.5, PE-Cy5.5 PerCP-eFluor710 |
585 BP 42 | PE, dsRed, dTomato, Alexa 546/555, Sytox Orange |
610 BP 20 | PE-CF594, Alexa 568/594, NovaFluor NY610, mCherry, PI, 7-aad |
660 BP 10 | PE-Cy5, NovaFluor NY660, mPlum, APC* |
780 BP 60 | PE-cy7 |
Spare filters | 610/20, 660/10, 712/25, 780/60. Can be swapped with any filter. No LP/SP required. |
*We have detected APC using the 660/10 filter off the 561 nm laser. |