Flow Cytometry & Bioimaging
The University of Surrey Bioimaging and Flow Cytometry Core Facility is situated within the Faculty of Health and Medical Science at the University of Surrey, Stag Hill Campus, Guildford.
We support the imaging and flow cytometry requirements of staff and students at the University of Surrey, as well as to outside academic and industrial institutions upon request. We have a wide research user base, with particular interests in cell biology, immunology, bacteriology, virology, cancer biology, and cell death.
We provide full training and any assistance required in flow cytometry and confocal microscopy for research students and staff. We also offer a cell sorting service. Please direct requests for equipment booking and training to Dr Rachel Butler, Facility Manager r.shrimpton@surrey.ac.uk.
| Flow Cytometry | Bioimaging | Useful Links | Contacts |
FLOW CYTOMETRY
The facility is equipped with the following flow cytometers and cell sorter:
1) BD Biosciences FACS Canto (2-laser, 6-colour), Windows XP computer with FACS DIVA v5.0
| Laser | Pos | Detector | Filter | Fluorochrome choices |
| 488nm | D | FITC | 530/30 | FITC, GFP, YFP, CFSE, CFDA, AlexaFluor488, calcein |
| C | PE | 585/42 | PE, PI | |
| B | PerCP* | 670LP | PI, 7AAD, PerCP, PerCP-Cy5.5, PE-Cy5** | |
| A | PE-Cy7 | 780/60 | PE-Cy7 | |
| 633nm | B | APC | 660/20 | APC, AlexaFluor647 |
| A | APC-Cy7 | 780/60 | APC-Cy7, APC-H7 |
Notes: * Detector also can be named PerCP-Cy5.5; ** PE-Cy5 and APC cannot be used together
2) BD Biosciences FACS Scan (1-laser, 3 colour), G4 PowerMac System 9.2, with CellQuest v3.3
| Laser | Detector | Filter | Fluorochrome choices |
| 488nm | FL-1 | 530/30 | FITC, GFP, YFP, CFSE, CFDA, AlexaFluor488, calcein |
| FL-2 | 585/42 | PE, PI | |
| FL-3 | 670LP | PI, 7AAD, PerCP, PerCP-Cy5.5, PE-Cy5 |
3) Beckman Coulter Epics XL (1-laser, 3 colour), Windows 98 PC with EXPO32
| Laser | Detector | Filter | Fluorochrome choices |
| 488nm | FL-1 | 525/30 | FITC, GFP, YFP, CFSE, CFDA, AlexaFluor488, calcein |
| FL-2 | 575/30 | PE, PI | |
FL-3 FL-4 | 620/30 675/30 | PI, PE-Texas Red 7AAD, PerCP, PE-Cy5 |
4) BD Biosciences FACS ARIA (3-laser, 9 colour), Windows XP computer with FACS DIVA v5.0
| Laser | Pos | Detector | Filter | Fluorochrome choices |
| 405nm | B | Violet 2 | 450/40 | DAPI, BD Horizon V450, Pacific Blue, Alexafluor405 |
| A | Violet 1 | 530/30 | BD Horizon V500, AmCyan, AlexaFluor430 | |
| 488nm | E | FITC | 530/30 | FITC, GFP, YFP, CFSE, CFDA, AlexaFluor488, calcein |
| D | PE | 575/26 585/42 | PE*, PI PE**, PI | |
| C | PE-Texas Red | 610/20 | PE-Texas Red, PI | |
| B | PerCP-Cy5.5 | 675/20 695/40 | PE-Cy5***, 7AAD PerCP, PerCP-Cy5.5 | |
| A | PE-Cy7 | 780/60 | PE-Cy7 | |
| 633nm | B | APC | 660/20 | APC, AlexaFluor647 |
| A | APC-Cy7 | 780/60 | APC-Cy7, APC-H7 |
Notes: *PE with PE-Texas Red; **PE without PE-Texas Red; ***PE-Cy5 and APC cannot be used together
BIOIMAGING
The facility is equipped with the following microscopes
Zeiss LSM510 VIS META Confocal Microscope (on an Axiovert 200M microscope
- 3 lasers and 6 laser lines:
Argon: 458nm, 477nm , 488nm ,514nm
HeNe1: 543nm
HeNe2: 633nm
- META spectral detector for spectral imaging
- Objective lenses:
Plan-Neofluar 10x/0.3, Plan-Neofluar 20x/0.5NA, Plan-Neofluar 40x/1.3NA oil DIC, C-Apochromat 63x/1.2 W corr, Plan-Apochromat 63x/1.4 oil DIC
- Motorised stage for automated multi area timelapse imaging
- Full environmental chamber for temperature control plus local CO2 delivery for atmospheric control
- Windows Vista computer with Zen 2009 confocal acquisition software
NEW – Second Confocal Microscope due for installation Summer 2013.
USEFUL LINKS
An excellent training video from BD Biosciences for learning about flow cytometry
http://www.bdbiosciences.com/services/training/itf_launch.jsp
A shorter but highly recommended training video from Molecular Probes
http://probes.invitrogen.com/resources/education/tutorials/4Intro_Flow/player.html
The Nikon online microscopy campus
http://www.microscopyu.com/articles/confocal/
The Zeiss online microscopy campus
http://zeiss-campus.magnet.fsu.edu/referencelibrary/laserconfocal.html
Check the excitation and emission spectra for your fluorochromes using this online tool from Invitrogen
http://www.invitrogen.com/site/us/en/home/Products-and-Services/Applications/Cell-Analysis/Labeling-Chemistry/Fluorescence-SpectraViewer.html
A table of the excitation and emission peaks of fluorochromes commonly used in microscopy and flow cytometry
http://flowcyt.salk.edu/fluo.html
Popular flow cytometry fluorochromes (BD Biosceinces)
http://www.bdbiosciences.com/documents/Multicolor_Fluorochrome_Guide.pdf
CONTACT DETAILS
Dr Rachel E Butler (Facility Manager)
Research Fellow in Bioimaging and Flow Cytometry
Department of Microbial and Cellular Sciences
Faculty of Health and Medical Sciences
University of Surrey
Guildford GU2 7XH
Email: r.shrimpton@surrey.ac.uk
Tel: +44 (0) 1483 686428
Mrs Rebecca Holland (Technical Support for Confocal)
Department of Biochemistry and Physiology
Faculty of Health and Medical Sciences
University of Surrey
Guildford GU2 7XH
Email: r.holland@surrey.ac.uk
Tel: +44 (0) 1483 686401
Professor Graham Stewart (Academic Lead for the Facility)
Professor in Molecular Bacteriology
Department of Microbial and Cellular Sciences
Faculty of Health and Medical Sciences
University of Surrey
Guildford GU2 7XH
Email: g.stewart@surrey.ac.uk
Tel: +44 (0) 1483 686423
