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  Institute of Cell and Molecular Science
Flow Cytometry Core Facility equipment, images used with permission by BD
  Flow Cytometry Core Facility
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Instruments available for use

There are currently six flow cytometers and a LED 4 colour epi-fluorescence microscope in the Core Facility.

The Flow Cytometry Core Facilities new in 2020 Cytek Aurora spectral unmixing 5 laser (Blue, Yellow-Green, UV, Red and Violet) with 96 well TC plate loader allows the detection of 40 fluorochromes over a 22 bit 6.5 log decade fluorescence scale and analyses 35,000 events/sec. The instrument has a full spectral array of PMTS from 370-890 nm with the Blue (488nm) laser detecting 14 parameters, Violet (40 nm) laser 16 parameters, Yellow-Green (561nm) laser 10 parameters, UV laser (360nm) 16 parameters and Red (640nm) laser 8 parameters. FSC and SSC (detected by Blue and Violet lasers). The instrument has FSC Express version 7 and Spectraflow software on a Windows 10 computer allows spectral unmixing of overlapping fluorochromes e.g. APC and AF647, BV421 & Pacific Blue.

The BD LSR II is an analyser with four lasers, a Blue (6 colours), Red (3 colours), UV (2 colours) and Violet lasers (3 colours) and thus capable of detecting 14 fluorescent signals e.g. FITC, PE, PE-Texas Red or Quantum Dot 605, PE-Cy5, PE-Cy5.5, PE-Cy7, APC, APC-Cy7, Alexa700, Pacific Blue or BV421, BV511 or Pacific Orange, BV605 and violet and green Indo-1 signals or BUV395 and BUV737. Fluorochrome list and Chromocyte pdf

The ACEA Novocyte is an analyser with three lasers, a Blue (5 colours), Red (2 colours) and a Violet lasers (6 colours) and thus capable of detecting 13 fluorescent signals e.g. FITC, PE, PE-Texas Red, PerCP or PE-Cy5 or PE-Cy5.5 or PerCP-Cy5.5, PE-Cy7, APC, APC-Cy7, Pacific Blue or BV421, BV511, BV570, BV605, BV650, BV785. The instrument is 24 bit technology allowing detection of 7.2 log decades of fluroescence and hence has no requirement to set PMT voltages. The instrument can perform absolute counts without the need for count beads, has a flow rate of up to 35,000 events/sec and is fitted with a TC plate & FACS tube reader, see pdf

The BD FACS Canto II is an analyser with three lasers, a Blue (4 colours), Red (2 colours) and a Violet lasers (2 colours) and thus capable of detecting 8 fluorescent signals e.g. FITC, PE, PE-Cy5.5, PE-Cy7, APC, APC-Cy7, Pacific Blue or BV421 and AmCyan or Pacific Orange or BV511. This instrument also has FACS Canto clinical software v2.1 suitable for clinical samples. Fluorochrome list and Chromocyte pdf

The Merck Millipore Muse is a small digital instrument fitted with an Green HeNe with 2 colour PMT detectors. These include PE/PI and PE-Cy5/7-AAD detectors. The 32 bit PMT technology removes the requirement to set voltages on the PMT by the operator in advance of acquiring the samples. The instrument can be used with range of purpose made kits, including annexin V, PI cell cycle and absolute number determinations without the need for the operator to set the compensation. There is a wide range of specially designed kits covering a wide range of biological processes, see link.

The BD FACSAria IIIu is a cell sorter with four lasers, an Blue laser (2 colours), Red , (3 colours) Yellow-Green (4 colours) and Violet diode (6 colours) and currently capable of detecting 15 fluorescent signals. Blue laser excites FITC, PerCP or PercP-Cy5.5: Yellow laser excites PE, PE-Texas Red or PE-CF95, PE-Cy5 or PE-Cy5.5, PE-Cy7: Red laser APC, Alexa700 or APC-Cy7: Violet laser BV421 or Pacific Blue, BV511 or Pacific Orange, BV605, BV650, BV711 and BV780. Chromocyte pdf

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, 15,000 or 10,000 cells per second for cells >10um via 70, 85um and 100um nozzle sizes at 70, 40 and 20 psi respectively.Fluorochrome list.

The Flow Cytometry Core Facility LED fluorescent microscope allows excitation at 4 wavelengths at 355, 425, 470 and 540 nm and emissions to be collected at 420, 450, 530 and 600nm, thus allowing the detection of DAPI, FITC & TRITC. This upright microscope is fitted with 4x, 10x, 40x and 100x objectives. The LED microscope is fitted with a digital 3 MP colour camera connected to a HP computer via a USB 2.0 cable. Images are collected via GXCAPTURE software (GT Vision).

 

 

 
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by Gary Warnes. © Queen Mary, University of London 2007
Institute of Cell and Molecular Science, The Blizard Building, Barts and The London School of Medicine and Dentistry, 4 Newark Street, London E1 2AT, UK Tel: +44 (0)20 7882 2483, Fax: +44 (0)20 7882 2200