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Flow Cytometry

 

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The Flow Cytometry Unit is located on floor 10 at Core Facility.

Flow Cytometry lab tel: 010-1033697

Staff

Florence Sjögren, tel: 010-1037718 (mini call 0740 556602)                 

Mikael Pihl: tel 010-1032292

 

Flow Cytometry Unit user group: Olle StålFlorence SjögrenJan-Ingvar Jönsson (chairman), Jan ErnerudhRosaura Casas, Anna Lundberg, Mikael Pihl and Robert Blomgran.

 

 

For documentational purposes, Flow Cytometry Core Facility would appreciate if all users alert the staff of any papers they publish utilizing the instruments at the facility. This helps us show our relevance to instances that determine our future funding.

Mentioning the core facility in i.e. the Materials and Methods section of your paper, for example when you specify what cytometer was used to acquire data or perform sorting, is also greatly appreciated. As an example, cells were acquired using a Gallios cytometer at the flow cytometry core facility of the faculty of health sciences, or similar. If appropriate, feel free to mention the contribution of the core facility in your acknowledgements - it would be greatly appreciated.

 


General Information

The Flow Cytometry Unit at the Faculty of Health Sciences is located on floor 10 at Core Facility. Flow Cytometry is used to measures physical and biological parameters of single cells in suspension. Cell size, granularity, protein expression on the surface and intracellularly and DNA-content are examples of properties that can be determined by FACS.

There are currently four Flow Cytometers available, two are capable of sorting individual cells into plates, 5ml tubes and 15ml tubes. All instruments are connected to a laser printer through a local network, but none of them have an internet connection.

Those available to answer questions and provide support, as well as keeping the instruments in working order are Florence Sjögren, tel 010-1037718 (mini call 0740 556602) and Mikael Pihl, tel 010-1034662. Florence's office is next to the cell sorters, and Mikael's office is outside KEF on floor 10.

As for every complex and sensitive instrument, there are rules for running our cytometers. A driver's license is also required to use any cytometer. A license is acquired through a user course, or through training by another experienced user. Contact Florence or Micke regarding this. "In house" user courses will be arranged as the need arises.

 

For more info on Flow Cytometry: www.invitrogen.com/site/us/en/home/support/Tutorials.html
Flow Analysis software that is available on the internet:
WinMDI from Scripps:
http://facs.scripps.edu/software.html
Flow Explorer from M Roederer: http://software.ronhoebe.com/indexFlowExplorer.html
MFI from U Mass: http://www.umass.edu/microbio/mfi/

Fluorochromes, filters etc:
http://flowcyt.salk.edu/fluo.html
http://www.biotium.com/
http://www.bdbiosciences.com/eu/research/multicolor/spectrum_viewer/index.jsp
http://www.bdbiosciences.com/documents/Multicolor_Fluorochrome_Guide.pdf#search=(fluorochromes)

 


Instruments


Cell sorters

ARIA

Sorteraren

The FACSAria is a high-speed cell sorter (ca 20000 cells/second) and is equipped with a UV-laser (355 nm) with detectors for 2 colors, a violet laser (405 nm) with detectors for 3 colors, an argon laser (488 nm) with detectors for 6 colors and a HeNe-laser (633 nm) with 3 detectors. 14 colors can be measured simultaneously. It is possible to sort 2-way and 4-way into tubes and to sort directly into plates. The instrument is operated using a PC running FACSDiva software. Filter configuration.
Fees to use:
Do it yourself: 120 sek/ hour.
Analysis consultation: 365 sek/ hour.
External user (non-LIU): 665 sek/ hour.

Reservation of instrument

 

ARIA III

Sorterare 2
The ARIA III is our latest sorter, it has the same capacity as the ARIA but has a different laser setup. Six detectors are available for the violet laser (405 nm), 3 for the blue (488 nm), 5 for the green (561 nm), and 3 for the red (632 nm) laser. A total of 17 fluorochromes can be detected simultaneously. The sorting capabilities are the same as on the ARIA. Filter configuration.

Additional filter info.


Fees to use:
Do it yourself: 120 sek/ hour.
Analysis consultation: 365 sek/ hour.
External user (non-LIU): 665 sek/ hour.

Reservation of instrument


Cytometers


Gallios

GalliosAn instrument for analysis from Beckman Coulter. Lasers available on this instruments are: violet laser (405 nm) 2 colors, argon laser (488 nm) 5 colors and  HeNe-laser (633 nm) 3 colors. The instrument is controlled with Navios software. Analysis of acquired data performed using Kaluza software on a separate PC.

Filter config:

Blue: FL1 550SP 525BP, FL2 595SP 575BP, FL3 655SP 620/30, FL4 730SP 695/30 - alt 675BP, FL5 755LP

Red: FL6 710SP 660BP, FL7 750SP 725/20, FL8 755LP

Violet: FL9 480SP 450/50, FL10 550/40

Fees to use:
Do it yourself: 120 sek/ hour.
Analysis consultation: 365 sek/ hour.
External user (non-LIU): 665 sek/ hour.

Reservation of instrument

 

 

 

 


Imaging Cytometer

ImageStreamx mkII

ISX

A flow cytometer that captures images of cells in 12 channels, equipped with blue and red lasers and able to acquire 9 fluorescence signals in addition to brightfield and darkfield. Three degrees of magnification are available (20X, 40X, 60X). The instrument is well suited for analysis of internalisation, apoptosis, translocation, nuclear localization and colocalization of two or more cellular components. In addition to fluorescense intensity data similar to that from a traditional cytometer, the ImageStream can produce numbers corresponding to roundness and other aspects of morphology for each individual cell. The instrument accepts sample columes between 20 and 200ul, recommended cell concentration is 20 million per ml. The machine only accepts Eppendorf tubes. 

 

Mer info

 


Mass Cytometer

CyTOF 2

CyTOF 2

The Flow Cytometry Unit is home to Sweden's first Mass Cytometer, a CyTOF 2 from DVS Sciences / Fluidigm, which is a part of the national resource for mass cytometry. Mass Cytometry substitutes Lanthanide isotopes for fluorochromes and separates them by mass using time-of-flight measurement. Problems with background signal due to autofluorescence and limitations due to spectral overlap are thus eliminated. There are approximately 40 different isotopes available that can be conjugated to antibodies, as well as live/dead staining options. It is however not possible to acquire morphological data like on a traditional cytometer.

Contact Jan-Ingvar Jönsson for further information.

 

 


 

Luminex 200

Luminex 200

A flow cytometty instrument aimed solely at analysing fluorescent latex beads. Allows quantification of several different soluble proteins simultaneously. It's also possible to analyze fluorescent magnetic beads. Controlled by xPONENT software.

Additional Information: http://www.luminexcorp.com/01_xMAPTechnology

Fees:
Do it yourself: 200 sek/plate.
Analysis consultation: 625 sek/run.
External user (non-LIU): 925 sek/run

Reservation of instrument

 

 

 

 


Analysis workstations, room 301

 Analysrummet

We provide three workstations for analysis of cytometry data in room 301 at Core Facility. A G5 mac for analysis of Cellquest Pro data, a PC for analysis using DIVA software and StarStation software for analysis of data acquired on the Luminex 200, and a PC running Kaluza software for analysis of any flow cytometry data.

Reservation of workstations

 

 

 

 

 

Additional Info

 

1. We gather for user meetings regularly, where users can put forward ideas or suggestions for improving the working environment and practices in the Flow Cytometry Unit. Information about these meetings will be spread via e-mail. We aim to achieve pleasant working conditions at the Flow Cytometry facility and an environment where scientists can support each other in their respective projects.
2. Suggestions for future investments in instrumentations, rules for using our equipment or needs for training or basic education in flow cytometry should be put forward to the user group, which consits of Olle Stål, Florence Sjögren, Jan-Ingvar Jönsson (chairman), Jan Ernerudh, Rosaura CasasAnna LundbergMikael Pihl och Robert Blomgran..


Page manager: mikael.pihl@liu.se
Last updated: 2019-03-13