Equipment access and operating procedures
To use the flow cytometry equipment, you must first undergo training with a Flow Cytometry staff member and be registered. You will then be added to the Flow Cytometry OMNI team on Microsoft Teams, where you can find up-to-date standard operating procedures.
Flow Cytometry OMNI on Microsoft Teams
To request equipment training and access, contact:
Email OMNIflow@otago.ac.nz
BC Cytoflex S
The Cytoflex is a semi-portable instrument and is available for use in teaching labs. Please contact the flow cytometry team if you would like to use this instrument for teaching purposes.
The Cytoflex S is a 3 laser, 9 detector benchtop system with modern generation technology giving much better sensitivity than other conventional cytometers. It also has great signal to noise resolution giving you a cleaner result.
NOTE: This machine is housed in the Pathology Department, Hercus Building, second floor, west wing lab (223).
| Laser | Bandpass | Fluorochrome |
|---|---|---|
| 488nm Blue | 525/40 | FITC, AF488, CFSE |
| 690/50 | PerCP, PerCP-Cy5.5, PI | |
| 561nm Yellow/Green |
610/20 | mCherry, PE-CF594 |
| 585/42 | PE | |
| 690/50 | PerCP, PerCP-Cy5.5, PI | |
| 780/60 | PC7, DRAQ7 | |
| 638nm Red | 660/10 | APC, AF647 |
| 712/25 | APC-A700, AF700, DRAQ5 | |
| 780/60 | APC-A750, APC-Cy7 |
BD FACSAria Fusion
The FACSAria is a 16 channel cytometer with cell sorting capability good for medium multicolour panels up to 12-14 colours.
This machine is contained within a Class II BSC and therefore can be used to work on unfixed material and maintain sterility. It also has a temperature controlled sample injection port and output. This can be heated or cooled as required but staff must be aware before use. This machine can perform, among other things, single cell sorting and fast 4-way sorting.
To use this machine the experiment must be designed with help from the facility staff, cells must be filtered through a 70um filter and staff will run the samples through the cytometer. There is also an added non-negotiable 30 minutes to each booking for machine set up and aseptic cleaning.
| Laser | Mirror | Filter | Fluorochrome |
|---|---|---|---|
| 488nm Blue |
502 | 530/30 | FITC, AF488, CFSE, Zombie Green, GFP |
| 655 | 695/40 | PerCP, PerCP-Cy5.5 | |
| 561nm Yellow/Green |
582/15 | PE, DsRed | |
| 600 | 610/20 | mCherry, PE-Cf594, PE-Texas red, Live/Dead red, PE-Dazzle, PI, Texas red | |
| 630 | 670/14 | PE-Cy5 | |
| 685 | 710/50 | PE-Cy5.5 | |
| 735 | 780/60 | PE-Cy7 | |
| 640nm Red |
670/30 | APC, AF647, Live/Dead Far red | |
| 690 | 730/45 | AF700 | |
| 755 | 780/60 | APC-Cy7, APC-H7, Live/Dead NIR | |
| 405nm Violet |
450/50 | Bv421, DAPI, Pacific Blue | |
| 505 | 525/50 | Bv510, Live/Dead Aqua, AmCyan, Live/Dead yellow | |
| 595 | 610/20 | Bv605 | |
| 630 | 660/20 | Bv650 | |
| 690 | 710/50 | Bv711 | |
| 750 | 780/60 | Bv786 |
BD FACSCanto II
The CANTO II is an 8 channel analyser cytometer, good for small multicolour panels up to 6 colours. This machine is contained within a Class II BSC and therefore can be used to work on unfixed material. Good cleaning still required!
| Wavelength | Mirror | Detector | Fluorochrome |
|---|---|---|---|
| 488nm Blue |
502 | 530/30 | FITC, AF488, Zombie Green, CFSE |
| 556 | 585/42 | PE, PI | |
| 655 | 670LP | PerCP, PerCP-Cy5.5 | |
| 735 | 780/60 | PE-Cy7 | |
| 633nm Red |
660/20 | APC, AF647 | |
| 735 | 780/60 | APC-Cy7, APC-H7 | |
| 405nm Violet |
450/50 | Bv421, Pacific Blue | |
| 502 | 510/50 | Bv510 |
BD LSRFortessa
The Fortessa is a 16 channel analyser cytometer, good for multicolour panels up to 10-12 colours.
| Laser | Mirror | Bandpass | Fluorochrome |
|---|---|---|---|
| 488nm Blue |
505 | 530/30 | FITC, GFP, AF488, CFSE, Zombie Green |
| 685 | 710/50 | PerCP-Cy5.5, PerCP, 7AAD | |
| 561nm Yellow/Green |
586/15 | PS-Dazzle 594, PE, AFS46 | |
| 600 | 610/20 | AFS68, AF594, PE-Cy5, Texas Red | |
| 635 | 670/30 | PE-Cy5.5, PE-Fire 640 | |
| 685 | 710/50 | PE-Cy5.5 | |
| 750 | 780/60 | PE-Cy7 | |
| 633nm Red |
670/14 | APC, AF647, APC-Cy5, DRAQ5 | |
| 690 | 730/45 | AF660, AF680, AF700 | |
| 735 | 780/60 | APC-Cy7, APCH7, APC-efluor 780, APC-fire 750, Live/Dead fixable Near IR, Zombie NIR | |
| 405nm Violet |
450/50 | Pac Blue, BV421, AF405, Zombie Violet, DAPI, CFP | |
| 505 | 525/50 | BV510, Zombie Aqua, Zombie Yellow, Live/Dead Fixable Aqua | |
| 600 | 610/20 | Bv605, eFluor 605, Superbright 600 | |
| 630 | 660/20 | Bv650, Superbright 645 | |
| 690 | 710/50 | Bv711, Bv750 | |
| 750 | 780/60 | Bv750, Bv785 |
Cytek Aurora
The Cytek Aurora is spectral analyser cytometer with immense potential. It detects the whole spectrum of emitted light. The machine here has 4 lasers, including ultraviolet, violet, blue and red. This modern machine is a game changer, enabling large panels that you could only previously see using mass cytometry. With good controls and planning, 25 to 30 colour panels are possible.
Due to the fundamental difference between standard cytometry and spectral cytometry, the flow unit offers comprehensive training in spectral cytometry panel design, machine/software use and big data analysis.
This machine has an automated sample loader attached for high throughput analysis. It can hold 96 well plates or up to 40x 5ml FACs tubes. It is also customisable. Please ask the OMNI flow team for more information.
There is no fluorochrome guide, as if the fluorochrome can be excited in this cytometer then it can be detected.