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ROOM 4027A - cell analysers

BD FACSymphony A5 (SORP)

BD FACSymphony A5 (SORP): (SN: R660964R5001) Manufactured in February 2018 and installed in March 2018. BD's flagship 50-parameter instrument.
This instrument uses 6 lasers (355nm, 405nm, 488nm, 561nm, 640nm & 785nm) to detect 30 parameters (FSC, SSC and 28 fluorescence channels), with advanced electronics adapted from the defence industry that allows event collection at up to 40,000 cells/sec. A software upgrade in the near future will extend this capability to 50-parameter acquisition. The instrument currently runs BD FACSDiva 8.0.2. The above analysis displays a 27- parameter panel allowing discrimination of 40+ different cell phenotypes.  
This instrument is connected to a BD FACS flow supply system, a simple to use fluidics cart for delivery of sheath fluid and collection of waste. It has an optional High Throughput Sampler (HTS) which allows sampling directly from 96- or 384- well plates.
 
 
 
BD LSRFortessa X20 (SORP) (Three instruments)
BD LSRFortessa X20 (SORP): These instruments use 5 lasers (355nm, 405nm, 488nm, 561nm & 640nm) to detect 20 parameters at up to 20,000 cells per second (FSC, SSC and 18 fluorescence channels). This instrument runs BD FACSDiva 8.0.1.
This instrument is connected to a BD FACS Flow Supply System, a simple to use fluidics cart for delivery of sheath fluid and collection of waste. It has an optional High Throughput Sampler (HTS) which allows sampling directly from 96- or 384- well plates.
All three BD LSRFortessa X-20 units are configured with the same laser and parameter filter configuration.
 
BD LSRFortessa X20 (SORP) #1: (SN: H658226M1001) Manufactured in December 2014, and installed in January 2015.
BD LSRFortessa X20 (SORP) #2 (Everest): (SN: H658226R1027) Manufactured in June 2016, and installed in July 2016.
BD LSRFortessa X20 (SORP) #3 (Mauna Kea): (SN: H658226R1053) Manufactured in April 2017, and installed in June 2017.
 
 
Merck-Millipore ImageStreamX MkII
Merck-Millipore AMNIS ImageStreamX MkII: (Serial No. 799-03300 / ISX 570) Manufactured in November 2018, and installed in March 2018.
The AMNIS ImageStreamX MkII is an instrument capable of performing imaging cytometry, an application combining the advantages of flow cytometry and microscopy in one unit. This 2-camera, 12- channel system allows collection of bright-field images of analysed cells, and is fitted with 405nm, 488nm, & 642nm lasers for fluorescence analysis of up to 10 fluorophores simultaneously (all data is recorded on to CCD camera, allowing imaging of fluorescence in cells giving localisation). The unit also has a 785nm laser for dark-field SSC analysis.
The instrument is equipped with the Multi-Mag option, allowing use of x20, x40 or x60 objectives and an EDF (Extended Depth of Field) module for enhanced spot count function.
Data acquisition (up to 5,000 cells per second) is conducted using INSPIRE software. Analysis is conducted offline with IDEAS software, a sophisticated image analysis software.
 
 
Mindray BC-5000 Vet Auto Haemotology Analyser
Mindray BC-5000 Vet Automated Haemotology Analyser: (Instance No. SX-6B000369) Manufactured in November 2016, acquired for tesing in January 2017 and installed in March 2017. 

This instrument uses electrical impedance, chemical staining and low-, medium- and wide- angle laser scatter to perform complete blood counts (CBC's) on whole blood reporting 23 parameters. This instrument can analyse samples at a throughput of 60 samples per hour, and uses a minimal sample volume (15 mL) in whole blood mode.

The Mindray enumerates absolute counts of platelets, erythrocytes and leukocytes (CBCs); being a 5-part diff unit it allows the differentiation of leukocytes into 5 subsets (Lymphocytes, Monocytes, Neutrophils, Eosinophils and Basophils). The instrument reports on numerous other haematological parameters (eg. haemoglobin levels, see below for full list).

The instrument has 15 pre programmed settings for defined species of animals (e.g. Mouse, Rat, Rabbit, Ferret, Pig, Cow, Sheep, Goat, Monkey (human), Llama, Horse, Dog, Cat, Panda, Red Panda) and can accommodate an additional 20 custom-programmed species settings. It can be used for absolute cell counts on a variety of other samples other than blood. 

Parameters measured include: WBC, Lym%, Mon%, Neu%, Bas%, Eos%, Lym#, Mon#, Neu#, Eos#, Bas#, RBC, HBG, HCT, MCV, MCH, MCHC, RDW-CV, RDW-SD, PLT, MPV, PDW, PCT.

Beckman Coulter CytoFLEX-S (Two instruments)

Beckman Coulter CytoFLEX-S: This instrument uses 4 lasers (405nm, 488nm, 561nm, 638nm) to detect FSC, SSC and up to 13 fluorescence channels at acquisition rates up to 25,000 cells/sec. The unit uses a peri-pump system to aspirate samples, hence a variety of tubes can be used to deliver samples (1.5mL microfuge tubes, 5mL polystyrene and polypropylene tubes and has an adaptor for 1.2mL microtitre tubes). Accommodates a variety of tubes as well as 96 well plates.

Both Beckman Coulter CytoFLEX-S instruments are configured with the same laser and parameter filter configuration. 

Beckman Coulter CytoFLEX-S (CytoFLEX #1): (Serial No. AW04010, Instance No. 21684277) This instrument was manufactured in January 2016. Purchased at the end of 2015 and installed in February 2016. This unit can be housed in a Class 2 Biohazard hood for analysis of Risk Category 2 pathogens. 

Beckman Coulter CytoFLEX-S (CytoFLEX #2 - GLYCOFLEX): (Serial No. AW03009, Instance No.21798277) This instrument was manufactured in January 2016. Purchased at the end of 2015 and installed February 2016. 

BD Accuri C6

BD Accuri C6: (SN:4969) Manufactured in September 2013, and installed in October 2014.

This is a simple to use 2 laser (488nm, 640nm), 6 parameter instrument (FSC,SSC and 4 fluorescence channels). It requires no PMT voltage adjustment, and fluorescence data is measured over a dynamic 7 decades of data.

The unit uses a peri-pump system to aspirate sample, hence a variety of tubes can be used to deliver sample (1.5mL microtitre tubes, 5mL polystyrene and polypropylene tubes).

The instrument uses BD Accuri C6 software for data acquisition. 

ROOM 4027A - CELL SORTERS

Beckman Coulter MoFlo Astrios EQ

Beckman Coulter MoFlo Astrios EQ: (Instance No. 3969736) Manufactured 2012, installed September 2012. EQ upgrade performed July 2015.

TRIfcs staff operate the Beckman Coulter Astrios EQ sorter within the flow core. This unit is a 7-laser (7-pinhole, 355nm, 405nm, 488nm, 532nm, 561nm, 592nm, 640nm) allowing the detection of up to 26 parameters (FSC, SSC & 24 fluorescence channels).

The Beckman Coulter Astrios EQ sorter is a jet-in-air based sorter capable of 6-way cell sorting in a number of different sort modes (independently operating on target sort populations), facilitating separation of up to 6 populations simultaneously. Sorted populations can be deposited into a variety of tubes. Indexed single cell sorting is also available into a variety of plate types including 96- and 384- well plates.

Temperature control allows for maintaining the environment of sample and sorted material. The instrument is housed within a Biological Safety Cabinet allowing for sorting of human cells, human cell lines and virally-transduced cells. 

BD FACSAria Fusion (SORP) (Two Instruments)

BD FACSAria Fusion SORP: TRI.fcs  staff operate two BD FACSAria Fusions. These units are 5-laser (5 pinhole, 355nm, 405nm, 488nm, 561nm and 640nm) to detect FSC,SSC and 18 fluorescence channels. The 5-laser configuration is near identical to the BD LSRFortessa X-20 units configuration (FSC, SSC, 18 fluorescence channels), allowing easy transfer of developed analysis panels to these instruments for sorting applications.

The FACSAria fusions are capable of 4-way cell sorting in a number of different sort modes, facilitating separation of up to 4 populations simultaneously. Sorted populations can be deposited into a variety of tubes. Indexed single cell sorting is also available into a variety of plate types including 96- and 384- well plates.

Temperature control allows for maintaining the environment of sample and sorted material. Both units are housed within Biological Safety Cabinets allowing for sorting of human cells, human cell lines and virally-transduced cells. 

These instruments utliise a cuvette-based flow cell allowing superior detection of some fluorophores compared to jet-in-air systems. 

BD FACSAria Fusion SORP (Fusion #1 5-Laser Leo): (SN: P658282740001) Manufactured July 2016, and installed September 2016. Upgraded to a 5-laser, 5-pinhole unit in June 2018. This instrument uses 5 lasers (5 pinhole, 355nm, 405nm, 488nm, 561nm, 640nm) to detect FSC, SSC and 18 fluorescence channels. 

BD FACSAria Fusion SORP (Fusion #2 5-Laser Len): (SN: P658282740001) Manufactured June 2017, and installed September 2017. This instrument uses 5 lasers (5 pinhole, 355nm, 405nm, 488nm, 561nm, 640nm) to detect FSC, SSC and 18 fluorescence channels.

Miltenyi autoMACS Pro Separator

Miltenyi autoMACS Pro Separator: (Serial No. 968). Manufactured in September 2011, and installed in December 2011.

This instrument uses MACS (Magnetic activated cell sorting), a bead based technology available from Miltenyi to isolated cells directly from whole blood, bone marrow or virtually any cell type from any species subject to antibody availability.

It can be used as an alternative or complement to droplet-based flow sorting. The unit is housed in a Bio-safety cabinet allowing sorting of human primary material.

Several modes of purification are available that revolve around the principles of positive- or negative- selection.

ROOM 4008 - SOFTWARE AND ANALYSIS WORKSTATIONS 

SOFTWARE

FlowJo v10.7.1: TRIfcs has 5 licenses of this software available on PCs within the analysis suite area licenced by dongles. This aftermarket/third party software can be used for analysing a wide range of flow cytometric data collected on a variety of acquisition software platforms, including but not restricted to; BD Accuri C6 software, BD FACSDiva, Kaluza, CytExpert2.0, CellQuest Pro, CXP and Summit. Data is imported into FlowJo in the .fcs file format.

With an intuitive interface, specialised analysis platforms and open-ended plugin architecture, FlowJo provides a rich analysis environment that is easy to use, versatile and extensible. 

The FlowJo workspace is a poweful statistical environment that is used for immunophenotyping, cell cycle, proliferation, kinetics studies, quantitative population comparison or plate screening assays to name a few.

BD FACSDiva 8.0.1: BD Digital Vantage acquisition software (FACSDiva) also allows analysis of flow cytometric data collected on acquisition/sorter units running FACSDiva in two ways. FACSDiva experiments can be exported as an 'Experiment' (.xml file format) and re-imported into FACSDiva for analysis, maintaining all previously generated plots, gates, hierachy information and statistical analyses. Alternative data in .fcs file format can be imported in the FACSDiva for analysis by normal means. 

BD FACSDiva 8.0.2: BD Digital Vantage acquisition software (FACSDiva) also allows analysis of flow cytometric data collected on acquisition/sorter units running FACSDiva in two ways. FACSDiva experiments can be exported as an 'Experiment' (.xml file format) and re-imported into FACSDiva for analysis, maintaining all previously generated plots, gates, hierachy information and statistical analyses. Alternative data in .fcs file format can be imported in the FACSDiva for analysis by normal means.

Kaluza for Gallios and Kaluza 1.5/2.0: This acquisition software (Kaluza G) & analysis software (Kaluza 1.5/2.0) are from Beckman Coulter. TRI.fcs has 5 copies of this software on analysis units licenced by a dongle-driven network-based site licence (if Kaluza 1.5 analysis software doesn't open, please check that all workstations with Kaluza in the analysis suite area are turned on). It offers an intuitive analysis interface for the analysis of flow cytometric data. 

ModFit: Software for analysis of cell cycle distribution/DNA content. It is also capable of analysis of cell division where data was generated by dilution of dye-labelled cells (e.g. CFSE, CTV,etc.)

IDEAS Software: Software for analysis of image-cytometry data generated on the Merck-Millipore AMNIS ImageStreamX MkII instrument.

BD Accuri C6 Software: Software for analysis of imaging-cytometry data generated on BD C6 Accuri. 

BD FCAP Array: BD software for the analysis of multiplex bead-based cytokine analysis kits.

ANALYSIS SUITE AREA WORKSTATIONS

All workstations are operating in the 64-bit format of their O/S except where noted.

Turbulent - FlowJo 10.7.1, Kaluza 2.1

Transitional - FlowJo 10.7.1, Kaluza 2.1, Modfit LJ Ver.6

Rotational - FlowJo 10.7.1, Kaluza 2.1

Viscous - IDEAS Image cytometry analysis software

Lava (32-bit) - BD FACSDiva 8.0.2

Laminar (32-bit) - FACSDiva 8.0.2

Compressible (32-bit) - FlowJo 10.6.1, Kaluza 1.5, BD FCAP Array, BD Accuri C6 Software.

NB. All data should be analysed from a local drive (i.e D: drive) and not across the server from data residing on T: drive. Please copy data from T:drive to the local drive (D: drive), conduct your analysis and transfer analysed data back to T: drive. Analysis workstations have an auto-delete function that removes data when it is 2 weeks old on a given analysis workstation. Deleted data cannot be recovered.