FLOW CYTOMETRY
The INGM FACS Facility offers to the researchers an efficient and reliable personalized flow cytometry service.
Equipment
- Two FACScantoII analyzers, equipped with 3 lasers (405, 488, and 633 nm) to analyze up to 10 parameters.
- A BD FACSymphony A5 analyzer, equipped with 5 lasers (355, 405, 488, 561, 633) to analyze up to 30 parameters.
- A FACSaria III SORP cell sorter, equipped with 5 lasers (355, 405, 488, 561 and 633 nm) to simultaneously analyze up to 20 parameters and to separate up to 4 cell subsets.
- A FACSaria III cell sorter, equipped with 4 lasers (405, 488, 561, and 633 nm) to simultaneously analyze up to 14 parameters and to separate up to 4 cell subsets.
- A new sorter MACSQuant® Tyto® , benchtop cell sorter equipped with 3 lasers, in BLS3 laboratory which allows for high speed, 10-parameter cell sorting.
- A Miltenyi Automacs Separator for high-speed magnetic cell sorting of multiple samples.
The major objectives of the facility are:
- Provide cell sorting service for INGM researchers and external collaborators.
- Perform sorting of samples from healthy donors or patients suffering from non-infectious diseases (inflammatory diseases, autoimmune diseases), cancer biopsies and organoids.
- Set up, maintain and develop flow cytometry and cell sorting technology for basic and advanced research.
- Directly provide sample acquisition and analysis in specific applications for INGM researchers and external collaborators.
- Provide on-site training and assistance to prepare biomedical researchers to the use of optical instrumentation and to introduce to advanced flow cytometry applications.
- Perform independent and collaborative research for the development of advanced flow cytometry protocols.
- Perform collaborative research with INGM groups where highly specific flow cytometry and cell biology competences are required.
Our areas of expertise are:
- Immuno-phenotyping and cell sorting of all immune cell subsets.
- Immuno-phenotyping of hematopoietic stem cell subsets.
- Immuno-phenotyping of mesenchymal stem cell.
- Cell sorting of viable cytokine producing cells.
- Multi-color extracellular and intracellular staining.
- DNA content analysis (single color or multi-color).
- Cell proliferation analysis.
- Apoptosis analysis.
- General cell sorting approaches for cell lines and transfected cells.
Activity
Our facility is equipped with two BD FACSCanto II analyzers, a BD FACSAria I cell sorter located in a BL3 room and dedicated to infected samples and a BD FACSaria III sorter dedicated to samples from healthy donors or patients suffering from non-infectious diseases (inflamatory diseases, autoimmune diseases, cancer ecc.).
Our areas of expertise are:
- Immuno-phenotyping and cell sorting of all immune cell subsets.
- Immuno-phenotyping of hematopoietic stem cell subsets.
- Immuno-phenotyping of mesenchymal stem cell.
- Cell sorting of viable cytokine producing cells.
- Multi-color intracellular staining.
- DNA content analysys (single color or multi-color).
- Cell proliferation analisys.
- Apoptosis.
- General cell sorting approaches for cell lines and transfected cells.
Team
| Nome / Name | Ruolo / Role | |
|---|---|---|
| Mariacristina Crosti | Facility Manager | crosti@ingm.org |
| Maria Lucia Sarnicola | Facility Assistant | sarnicola@ingm.org |
Selected publications
- Recognition of viral and self-antigens by TH1 and TH1/TH17 central memory cells in patients with multiple sclerosis reveals distinct roles in immune surveillance and relapses.
Paroni M, Maltese V, De Simone M, Ranzani V, Larghi P, Fenoglio C, Pietroboni AM, De Riz MA, Crosti MC, Maglie S, Moro M, Caprioli F, Rossi R, Rossetti G, Galimberti D, Pagani M, Scarpini E, Abrignani S, Geginat J.
J Allergy Clin Immunol (2017) 140:797-808 - Extracellular MicroRNA Signature of Human Helper T Cell Subsets in Health and Autoimmunity.
Torri A, Carpi D, Bulgheroni E, Crosti MC, Moro M, Gruarin P, Rossi RL, Rossetti G, Di Vizio D, Hoxha M, Bollati V, Gagliani C, Tacchetti C, Paroni M, Geginat J, Corti L, Venegoni L, Berti E, Pagani M, Matarese G, Abrignani S, de Candia P.
J Biol Chem (2017) 292:2903-2915 - Transcriptional Landscape of Human Tissue Lymphocytes Unveils Uniqueness of Tumor-Infiltrating T Regulatory Cells.
De Simone M, Arrigoni A, Rossetti G, Gruarin P, Ranzani V, Politano C, Bonnal RJ, Provasi E, Sarnicola ML, Panzeri I, Moro M, Crosti M, Mazzara S, Vaira V, Bosari S, Palleschi A, Santambrogio L, Bovo G, Zucchini N, Totis M, Gianotti L, Cesana G, Perego RA, Maroni N, Pisani Ceretti A, Opocher E, De Francesco R, Geginat J, Stunnenberg HG, Abrignani S, Pagani M.
Immunity (2016) 45:1135-1147 - IL-10-producing forkhead box protein 3-negative regulatory T cells inhibit B-cell responses and are involved in systemic lupus erythematosus.
Facciotti F, Gagliani N, Haringer B, Alfen JS, Penatti A, Maglie S, Paroni M, Iseppon A, Moro M, Crosti MC, Stolzel K, Romagnani C, Moroni G, Ingegnoli F, Torretta S, Pignataro L, Annoni A, Russo F, Pagani M, Abrignani S, Meroni P, Flavell R, Geginat J.
J Allergy Clin Immunol (2016) 137:318-321 e5 - Dissection of the cord blood stromal component reveals predictive parameters for culture outcome.
Barilani M, Lavazza C, Vigano M, Montemurro T, Boldrin V, Parazzi V, Montelatici E, Crosti M, Moro M, Giordano R, Lazzari L.
Stem Cells Dev (2015) 24:104-14 - Reference proteome of highly purified human Th1 cells reveals strong effects on metabolism and protein ubiquitination upon differentiation.
Pagani M, Rockstroh M, Schuster M, Rossetti G, Moro M, Crosti M, Tomm JM.
Proteomics (2015) 15:3644-7 - IL-21 is a central memory T cell-associated cytokine that inhibits the generation of pathogenic Th1/17 effector cells.
Kastirr I, Maglie S, Paroni M, Alfen JS, Nizzoli G, Sugliano E, Crosti MC, Moro M, Steckel B, Steinfelder S, Stolzel K, Romagnani C, Botti F, Caprioli F, Pagani M, Abrignani S, Geginat J.
J Immunol (2014) 193:3322-31 - Human CD1c+ dendritic cells secrete high levels of IL-12 and potently prime cytotoxic T-cell responses.
Nizzoli G, Krietsch J, Weick A, Steinfelder S, Facciotti F, Gruarin P, Bianco A, Steckel B, Moro M, Crosti M, Romagnani C, Stolzel K, Torretta S, Pignataro L, Scheibenbogen C, Neddermann P, De Francesco R, Abrignani S, Geginat J.
Blood (2013) 122:932-42 - Identification of new hematopoietic cell subsets with a polyclonal antibody library specific for neglected proteins.
Moro M, Crosti M, Creo P, Gallina P, Curti S, Sugliano E, Scavelli R, Cattaneo D, Canidio E, Marconi M, Rebulla P, Sarmientos P, Viale G, Pagani M, Abrignani S.
PLoS One (2012) 7:e34395 - Distinct microRNA signatures in human lymphocyte subsets and enforcement of the naive state in CD4+ T cells by the microRNA miR-125b.
Rossi RL, Rossetti G, Wenandy L, Curti S, Ripamonti A, Bonnal RJ, Birolo RS, Moro M, Crosti MC, Gruarin P, Maglie S, Marabita F, Mascheroni D, Parente V, Comelli M, Trabucchi E, De Francesco R, Geginat J, Abrignani S, Pagani M.
Nat Immunol (2011) 12:796-803
INGM RESEARCH IS SUPPORTED BY
