Instrumentation

There are currently six flow cytometers and a Luminex LX100, and Mesoscale MSD in the core facility.

BECTON DICKINSON (BD) INSTRUMENTS

The facility has four BD instruments.

BD FORTESSA

The BD FORTESSA is an analyser with five lasers,

- Argon 488nm laser (6 colours),

- Red HeNe 640nm (2 colours),

- UV  355nm laser (2 colours) and

- Violet diode 405nm laser  (3 colours) and

- Yellow-Green 561nm laser (4 colours).

Thus capable of detecting 18 fluorescent signals e.g. FITC, PE, PE-Texas Red or Quantum Dot 605, PE-Cy5, PE-Cy5.5, PE-Cy7, APC, APC-Cy7, Pacific Blue, Pacific Orange, Quantum Dot 605 and violet and green Indo-1 signals. Fluorochrome list.

BD LSRII

The BD LSRII is an analyser with four lasers, an argon laser (6 colours), Red HeNe (2 colours), UV laser (2 colours) and violet diode (3 colours) and thus capable of detecting 13 fluorescent signals e.g. FITC, PE, PE-Texas Red or Quantum Dot 605, PE-Cy5, PE-Cy5.5, PE-Cy7, APC, APC-Cy7, Pacific Blue, Pacific Orange, Quantum Dot 605 and violet and green Indo-1 signals. Fluorochrome list.

BD FACS CANTOII HTS

The BD FACS CantoII HTS is an analyser with three lasers,an argon laser (4 colours), Red HeNe (2 colours) and a violet laser (2 colours) and thus capable of detecting 8 fluorescent signals e.g. FITC, PE, PE-Cy5.5, PE-Cy7, APC, APC-Cy7, Pacific Blue and Pacific Orange. This instrument also has FACS Canto clinical software v2.1 suitable for clinical samples. Fluorochrome list.

BD FACS ARIAII

The BD FACS Aria II is an analyser and cell sorter with three lasers, an argon laser (6 colours), Red HeNe (3 colours) and violet diode (3 colours) and capable of detecting 12 fluorescent signals e.g. FITC, PE, PE-Texas Red or Quantum Dot 605, PE-Cy5, PE-Cy5.5, PE-Cy7, APC, APC-Cy5.5, APC-Cy7, Pacific Blue, Pacific Orange, Quantum Dot 605. The FACSAria can perform 2 and 4 way sorting as well as automatic cell deposition for sorting onto TC plates or slides. Sort rates are 20,000 cells per second for lymphocytes and 10,000 cells per second for cells >10um via 70 and 100um nozzle sizes at 70 and 20 psi respectively. A 130um nozzle is now available which can sort cells >40um at 5,000 cells per second at 12 psi. Fluorochrome list.

DAKO CyanADP INSTRUMENTS

There are two CyAnADP's within this facility. These are compact benchtop flow cytometers from Beckman Coulter. Our CyAn's are  equipped with a 488nm blue laser, a 635 nm red diode and 405nm Violet laser.

The blue 488nm laser allows detection of seven parameters, two of which are forward and side scatters, and five enable simultaneous detection of fluorescence of 5 standard fluorochromes that emit at the following spectrum ranges:

- 530/40 (GFP, YFP, Alexa 488, FITC, etc...),

- 575/25 (PE, PI, etc...),

- 613/20 (mRFP, mCherry, PE-Texas-Red, PI, etc..),

- 680/30 (PECy5, PerCP, PerCPCy5.5, PI, 7AAD, etc...),

- 750LP (PECy7).

With its red diode, CyAn can detect fluorochromes that are excited at 635nm and emit at the following spectrum ranges:

- 665/20 (APC and its conjugates, Alexa 633, Alexa 647, ToPro-3, Draq5, etc...)

- 750LP (APCCy7, APC-H7, etc...)

With its violet laser, CyAn can detect fluorochromes that are excited at 405nm and emit at the following spectrum ranges:

- 450/50 (Alexa fluor 405, BD Horizon 450, BV421, DAPI, Pacific Blue, Sytox Blue, etc....)

- 530/40 (AmCyan, BD Horizon 500, Krome Orange; LIVE/DEAD Fixable Aqua Pacific Orange, etc...)

 Fluorochrome list