FACS
The FACS Facility at BRIC is a state-of-the-art multi-colour flow cytometry facilitymulti-colour flow cytometry facility, and we serve all research groups at BRIC, Finsen Lab, Bartholin Institute and others.
The FACS Facility at BRIC is a state-of-the-art multi-colour flow cytometry facility with Two Aria III, one FACS Melody, one LSR Fortesa X20, FACS Celesta and Accuri.
In addition to identification and isolation of various cell populations using antibodies, flowcytometry can analyze:
- Cell proliferation
- Cell physiological properties such as Calcium flux and pH
- DNA content and integrity, DNA base ratio
- Transfection markers such as GFP, mCherry, YFP, CFP, BFP, RFP and tdTomato
- Quantification of cytokines, gene expressions, microRNA, mRNA
- ROS assay
- Sulfhydrul groups/gluthathione quantification
- Receptor occupancy
- Phagocytosis
- Flowcytometry Protein Interaction Assay (FCPIA)
The Facility is located in lab 2-3-04 on the third floor and serves all research groups at BRIC, Finsen Lab, Bartholin Institute and others.
- Cell sorting into different plates (384, 96, 24, 6 well, PCR Plate, Index sorting), tubes and slides.
- FACS analysis
- Advice and supervision of users for experiments, trouble shooting and data analysis
- Multicolour panel design
- FACS training courses, individual one-to-one training (Only BRIC users)
- Involve in developing new protocols and techniques
- Assistance with Flowjo (An analysis software for data acquired in Flow cytometers).
Please contact:
Rajesh Somasundaram,
rajesh.somasundaram@bric.ku.dk
Phone: +45 3533 3177
For using the sorters or analyzers we have the following prices:
Do it yourself: 150 DKK/hour
Academic: 500 DKK/hour
Industrial: 1400 DKK/hour
Consultation: 500 DKK/hour
Instruments
Cell sorters
FACS AriaIII with 3 lasers and 17-parameter high speed cell sorting
15-color New Aria III configuration
Analysers
PEER-REVIEWED PUBLICATIONS
Ahsberg J, Xiao P, Okuyama K, Somasundaram R, Strid T, Qian H, Sigvardsson M. Progression of progenitor B cell leukemia is associated with alterations of the bone marrow micro-environment. Haematologica. 2019 Jul 11. pii: haematol.2018.214031. doi: 10.3324/haematol.2018.214031
Okuyama K, Strid T, Kuruvilla J, Somasundaram R, Cristobal S, Smith E, Prasad M, Fioretos T, Lilljebjörn H, Soneji S, Lang S, Ungerbäck J, Sigvardsson M. PAX5 is part of a functional transcription factor network targeted in lymphoid leukemia. PLoS Genet. 2019 Aug 5;15(8):e1008280. doi: 10.1371/journal.pgen.1008280. eCollection 2019 Aug
Somasundaram R, Ahsberg J, Okuyama K, Ungerback J, Lilljebjorn H, Fioretos T, Strid T, Sigvardsson M. 2016. Clonal conversion of B lymphoid leukemia reveals cross-lineage transferof malignant states. Genes Dev 30: 2486-99
Jensen CT, Åhsberg J, Sommarin MNE, Strid T, Somasundaram R, Okuyama K, Ungerbäck J, Kupari J, Airaksinen MS, Lang S, Bryder D, Soneji S, Karlsson G, Sigvardsson M. Dissection of progenitor compartments resolves developmental trajectories in Blymphopoiesis. J Exp Med. 2018 Jul 2;215(7):1947-1963
Hu Y, Zhang Z, Kashiwagi M, Yoshida T, Joshi I, Jena N, Somasundaram R, Emmanuel AO, Sigvardsson M, Fitamant J, El-Bardeesy N, Gounari F, Van Etten RA, Georgopoulos K. 2016. Superenhancer reprogramming drives a B-cell-epithelial transition and high-risk leukemia. Genes Dev 30: 1971-90
Somasundaram R, Fernandes S, Deuring JJ, de Haar C, Kuipers EJ, Vogelaar L, Middleton FA, van der Woude CJ, Peppelenbosch MP, Kerr WG, Fuhler GM. 2017. Analysis of SHIP1 expression and activity in Crohn's disease patients. PLoS One 12: e0182308
Jensen CT, Lang S, Somasundaram R, Soneji S, Sigvardsson M. 2016. Identification of Stage-Specific Surface Markers in Early B Cell Development Provides Novel Tools for Identification of Progenitor Populations. J Immunol 197: 1937-44
Ungerback J, Ahsberg J, Strid T, Somasundaram R, Sigvardsson M. 2015. Combined heterozygous loss of Ebf1 and Pax5 allows for T-lineage conversion of B cell progenitors. J Exp Med 212: 1109-23
Prasad MA, Ungerback J, Ahsberg J, Somasundaram R, Strid T, Larsson M, Mansson R, DePaepe A, Lilljebjorn H, Fioretos T, Hagman J, Sigvardsson M. 2015. Ebf1 heterozygosity resultsin increased DNA damage in pro-B cells and their synergistic transformation by Pax5 haploinsufficiency. Blood 125: 4052-9
Parikh K*, Zhou L*, Somasundaram R*, Fuhler GM, Deuring JJ, Blokzijl T, Regeling A, Kuipers EJ, Weersma RK, Nuij VJ, Alves M, Vogelaar L, Visser L, de Haar C, KrishnadathKK, van der Woude CJ, Dijkstra G, Faber KN, Peppelenbosch MP. 2014. Suppression of p21Rac signaling and increased innate immunity mediate remission in Crohn's disease. Sci Transl Med 6: 233ra53 * Equal contribution
Somasundaram R*, Nuij VJ*, van der Woude CJ, Kuipers EJ, Peppelenbosch MP, Fuhler GM. 2013. Peripheral neutrophil functions and cell signalling in Crohn`s disease. PLoS One 8: e84521. * Equal contribution
Zhou L*, Somasundaram R*, Nederhof RF, Dijkstra G, Faber KN, Peppelenbosch MP, Fuhler GM. Impact of human granulocyte and monocyte isolation procedures on functional studies. Clin Vaccine Immunol 19: 1065-74 *Equal contribution
Yuvaraj S, Al-Lahham SH, Somasundaram R, Figaroa PA, Peppelenbosch MP, Bos NA. 2012. E. coli-produced BMP-2 as a chemopreventive strategy for colon cancer: a proof-of-concept study. Gastroenterol Res Pract 2012: 895462
Somasundaram R, Deuring JJ, van der Woude CJ, Peppelenbosch MP, Fuhler GM. 2012. Linking risk conferring mutations in NCF4 to functional consequences in Crohn's disease. Gut 61: 1097; author reply -8
Hendricks J, Terpstra P, Dammers PM, Somasundaram R, Visser A, Stoel M, Bos NA, Kroese FG. 2010. Organization of the variable region of the immunoglobulin heavy-chain gene locusof the rat. Immunogenetics 62: 479-86
Fuhler GM, Baanstra M, Chesik D, Somasundaram R, Seckinger A, Hose D, Peppelenbosch MP, Bos NA. 2010. Bone marrow stromal cell interaction reduces syndecan-1 expression and induces kinomic changes in myeloma cells. Exp Cell Res 316: 1816-28
Review article
Somasundaram R, Prasad MA, Ungerback J, Sigvardsson M. 2015. Transcription factor networks in B-cell differentiation link development to acute lymphoid leukemia. Blood 126: 144-52
Regeling A*, Somasundaram R*, de Haar C, van der Woude CJ, Braat H, Peppelenbosch MP. 2010. Role of defective autophagia and the intestinal flora in Crohn disease. Self Nonself 1: 3237 * Indicates equal contribution
Somasundaram R, Strid T, Sigvardsson M. Cellular plasticity in B-Cell leukemia. Cell Cycle. 2017 Mar 19;16(6):495–496.
Theory about Flowcytometry
Flow Cytometry – A Basic Introduction
Michael G. Ormerod
BD Biosciences – Flow Cytometry Web-Based Training
Bio-Rad - Introduction to Flow Cytometry – Basics Guide
Guidelines
Guidelines for the use of flow cytometry and cell sorting in immunological studies (2nd ed)
Cossarizza et al, (2019)
Bible of Flowcytometry
Practical Flow Cytometry
Howard Shapiro
Spectral viewers for panel design
Spectral viewer – Thermo Fisher
Spectral viewer - BD Biosciences
Flourescence spectra Analyser - Biolegend
Flourofinder – Search and design platform
Learning links
Why antibody titration is important?
Contact
Rajesh Somasundaram
rajesh.somasundaram@bric.ku.dk
Phone: +45 35 33 31 77
BRIC - Biotech Research & Innovation Centre
Ole Maaløesvej 5
DK-2200 Copenhagen
Lab 2-3-04, third floor
Office 2-3-19