Spectral flow cytometry unit

The INGM Spectral Flow Cytometry Unit provides state-of-the-art spectral flow cytometry services for high-dimensional single-cell analysis, supporting both basic and translational research. The Unit offers scientific support throughout the entire experimental workflow, from experimental design and spectral panel optimization to computational data analysis and biological interpretation. The Unit has extensive expertise in the immunophenotyping of diverse cell populations, the design of complex spectral flow cytometry panels, and the identification of cell-specific autofluorescence signatures. It also has specialized expertise in extracellular vesicle characterization and provides scientific support for the development and optimization of innovative flow cytometry methodologies.

Equipment

Cytek Aurora Evo Spectral Flow Cytometer

The Spectral Flow Cytometry Unit is equipped with a Cytek Aurora Evo featuring five lasers (355, 405, 488, 561 and 640 nm) and full-spectrum detection technology. The system collects the complete emission spectrum of each fluorochrome, enabling high-dimensional analysis of more than 40 fluorescent parameters simultaneously while improving resolution of highly complex multicolor panels.

The instrument supports:

  • High-dimensional immunophenotyping
  • Full spectral unmixing
  • Multiple Autofluorescence extraction
  • High-sensitivity detection of rare populations
  • SPF module for enhanced resolution of small particles

Areas of technological expertise

  • High-dimensional immune profiling.
  • Spectral panel design and optimization.
  • Flow cytometry-based metabolic assays.
  • Assessment of fluorochrome spectral behavior.
  • Instrument optimization for the detection and characterization of nanoparticles, including extracellular vesicles.
  • Advanced computational data analysis using UMAP, FlowSOM, Phenograph, and related algorithms.

Area of scientific expertise

The Spectral Flow Cytometry Unit provides not only technical services but also scientific support for collaborative research projects.

The Unit has extensive expertise in immunology, with a focus on tumor immunology, cardiovascular diseases, immune monitoring, and functional immune cell characterization. It supports both human studies and preclinical research using murine models. The unit has also recognized expertise in extracellular vesicles as circulating biomarkers, including assay development, methodological optimization, and the application of spectral flow cytometry to liquid biopsy research.

Services

  • Support in experimental design
  • Panel design and optimization
  • Antibody and fluorochrome selection
  • Spectral unmixing
  • Computational analysis
  • Training and technical support

Team

Nome / Name Ruolo / Role Email
Achille Anselmo PhD Head anselmo@ingm.org
Daniele Pastorelli PhD Staff Scientist pastorelli@ingm.org

Selected publications

  • Vitiello PP, Rousseau B, Chilà R, Battuello P, Amodio V, Battaglieri V, Grasso G, Scardellato S, Anselmo A, Clemente F, Rospo G, Lamba S, Bartolini A, Pisati F, Tripodo C, Congiusta N, Russo M, Crisafulli G, Di Nicolantonio F, Germano G, Diaz LA Jr, Bardelli A. Cisplatin and temozolomide combinatorial treatment triggers hypermutability and immune surveillance in experimental cancer models. Cancer Cell. 2025 Jul 14;43(7)
  • Boselli D, Clemente F, Di Terlizzi S, Pagiatakis C, Papa L, Del Zotto G, Villa C, Ramirez GA, Maugeri N, Manfredi AA, Anselmo A. Unravelling Plasma Extracellular Vesicle Diversity With Optimised Spectral Flow Cytometry. J Extracell Biol. 2025 Apr 25;4(4).
  • Faccani C, Rotta G, Clemente F, Fedeli M, Abbati D, Manfredi F, Potenza A, Anselmo A, Pedica F, Fiorentini G, Villa C, Protti MP, Doglioni C, Aldrighetti L, Bonini C, Casorati G, Dellabona P, De Lalla C. Workflow for high-dimensional flow cytometry analysis of T cells from tumor metastases. Life Sci Alliance. 2022 Jun 3;5(10)
  • Anselmo A, Frank D, Papa L, Viviani Anselmi C, Di Pasquale E, Mazzola M, Panico C, Clemente F, Soldani C, Pagiatakis C, Hinkel R, Thalmann R, Kozlik-Feldmann R, Miragoli M, Carullo P, Vacchiano M, Chaves-Sanjuan A, Santo N, Losi MA, Ferrari MC, Puca AA, Christiansen V, Seoudy H, Freitag-Wolf S, Frey N, Dempfle A, Mercola M, Esposito G, Briguori C, Kupatt C, Condorelli G. Myocardial hypoxic stress mediates functional cardiac extracellular vesicle release. Eur Heart J. 2021 Jul 21;42(28):2780-2792.
  • Donadon M, Torzilli G, Cortese N, Soldani C, Di Tommaso L, Franceschini B, Carriero R, Barbagallo M, Rigamonti A, Anselmo A, Colombo FS, Maggi G, Lleo A, Cibella J, Peano C, Kunderfranco P, Roncalli M, Mantovani A, Marchesi F. Macrophage morphology correlates with single-cell diversity and prognosis in colorectal liver metastasis. J Exp Med. 2020 Nov 2;217(11):e20191847.
  • Galletti G, De Simone G, Mazza EMC, Puccio S, Mezzanotte C, Bi TM, Davydov AN, Metsger M, Scamardella E, Alvisi G, De Paoli F, Zanon V, Scarpa A, Camisa B, Colombo FS, Anselmo A, Peano C, Polletti S, Mavilio D, Gattinoni L, Boi SK, Youngblood BA, Jones RE, Baird DM, Gostick E, Llewellyn-Lacey S, Ladell K, Price DA, Chudakov DM, Newell EW, Casucci M, Lugli E. Two subsets of stem-like CD8 + memory T cell progenitors with distinct fate commitments in humans. Nat Immunol. 2020 Dec;21(12):1552-1562.
  • Cossarizza A, Chang HD, Radbruch A, Acs A, Adam D, Adam-Klages S, Agace W, Aghaeepour N, Akdis M, Allez M, Almeida LN, Alvisi G, Anderson G, Andrä I, Annunziato F, Anselmo A, et al. Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition). Eur J Immunol. 2019 Oct;49(10):1457-1973.
  • Veronica Larcher, Paolo Kunderfranco, Marco Vacchiano, Pierluigi Carullo, Marco Erreni, Irene Salamon, Federico Simone Colombo, Enrico Lugli, Marta Mazzola, Achille Anselmo* and Gianluigi Condorelli*. An autofluorescence-based method for the isolation of highly purified ventricular cardiomyocytes. Cardiovasc Res. 2018 Mar 1;114(3):409-416. *The authors are equally contributed
  • Anselmo A, Riva F, Gentile S, Soldani C, Barbagallo M, Mazzon C, Feruglio F, Polentarutti N, Somma P, Carullo P, Angelini C, Bacci M, Mendolicchio GL, Voza A, Nebuloni M, Mantovani A, Garlanda C. Expression and function of IL-1R8 (TIR8/SIGIRR): a regulatory member of the IL-1 receptor family in platelets. Cardiovasc Res. 2016; 111, 373–384
  • Anselmo A, Mazzon C, Borroni EM, Bonecchi R, Graham GJ, Locati M. Flow cytometry applications for the analysis of chemokine receptor expression and function. Cytometry A. 2014 Apr;85(4):292-301

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