University of Dundee

Research Technologies

Analysers

LSR Fortessa (Becton Dickinson): 

The LSR Fortessa is our custom specification, high end digital instrument and is located in the cell sorting room on the first floor in the MSI/WTB link corridor.

It is equipped with five laser light sources; 488nm, 640nm, 355nm, 405nm and 561nm, and up to 15 colours can be detected at any one time. The cytometer runs using PC based DIVA software and is equiped with full cross-beam compensation and pulse processing cabailities on all parameters.

FACS Canto (Becton Dickinson): 

The FACS Canto is also located in the Division of Cell Signalling and Immunology on the second floor of the Wellcome Trust Biocentre. Like the LSR Fortessa, the FACS Canto is a digital system and runs using the PC based DIVA software.

It has three excitation light sources: 405nm and 488nm solid state lasers and a 633nm laser. From these light sources, up to eight colours can be detected: 450nm and 510nm from the 405nm laser; 525nm, 575nm, 678nm and 785nm from the 488nm laser; and 660nm and 785nm from the 633nm laser. The cytometer is able to perform pulse processing on all parameters and also allows full inter-beam compensation, either during or post data acquisition.

FACS Verse (Becton Dickinson):

The FACS Verse is located in the Division of Cell Signalling and Immunology on the second floor of the Wellcome Trust Biocentre.

It has three laser exciation sources: 488nm, 635nm and 405nm, and is able to detect eight fluorescence parameters: four from 488nm, two from 635nm and two from 405nm. Optical filters can easily be changed to allow optimisation of detection of many different fluorophores.

The Verse is equiped with a volumetric flow sensor, which allows accurate counting of cells while performing analyses. It also has a universal loader which enables automated sample uptake from a number of different vessles, including including 384-well and 96 well plates.

Cell Sorters

Influx (Becton Dickinson):

The Influx is equiped with three laser excitation sources: 488nm, 635nm and 405nm, and is able to detect eleven fluorescence parameters simultanously. Its modular design means that additional excitation sources and detectors can easily be added as and when required.

The instrument is able to sort cells at a rate of up to 25,000 cells per second (depending on cell type!) and up to four different populations can be collected at any one time. Cells can be depositied into a variety of different vessles, including 96-well plates for single cell cloning, and temperature controlled sample and collection chambers preserve sample quality.

Only cells/samples designated as class 1 biohazardous material can be sorted on the Influx. Unlike the analysers within the facility, the cell sorter can only be operated by facility staff.

For more detailed information on what is required to successfully sort cells, see Techniques.

SH800 (Sony):

The Sony SH800 is the latest addition to our instrument portfolio. It is equipped with four laser excitation sources: 488nm, 405nm, 640nm and 561nm. The addition of the 561nm laser enables us to sort cells expressing fluorescent protiens such as mCherry and mKate. The sorter is able to detect up to six different fluorescent parameters and the changeable filter sets allows modification of the detectors to any combination of colours from any of the four lasers.

The sorter is housed in a class 2 biosafety cabinet, enabling us to sort samples derived from primary human tissue and those transfected with non-replicating human viruses.

The instrument is able to sort cells at a rate of up to 10,000 cells per second (depending on cell type!) and up to two different populations can be collected at any one time. Like the Influx, cells can be depositied into a variety of different vessles, including 384-well and 96-well plates for single cell cloning, and temperature controlled sample and collection chambers preserve sample quality.

Data Analysis

The Flow Cytometry and Cell Sorting Facility provides a Mac computer equipped with FlowJo software designated solely for data analysis. However, many groups who perform a lot of flow cytometry choose to purchase their own software to enable data analysis on their own computers. The most commomly used software in use is FlowJo, which is available for use on both Macs and PCs. Numerous other software packages are available from various sources.