Flow Cytometry

EQUIPMENTS

Cell Analysers

• FACSDiscover A8: 5 lasers – 355 nm, 405 nm, 488 nm, 561 nm, and 637 nm; enables high-dimensional spectral analysis of up to 80 parameters, featuring CellView technology for simultaneous real-time cellular imaging
• BD FACSCanto II: 3 lasers – 405nm, 488nm and 633nm; multicolor analysis of up to 8 parameters
• FACSCalibur: 2 lasers – 488nm and 633nm; multicolor analysis of up to 4 parameters

Cell Sorter

• BD FACSAria III: 4 lasers 405nm, 488nm, 561 and 640nm; cell sorter and multicolor analysis of up to 16 parameters
• Equipment fully integrated Class II Type A2 biosafety cabinet

SERVICES

• Maintenance and Quality Control of Scientific Equipment
  The Flow Cytometry Facility implements rigorous maintenance and quality control protocols, ensuring the optimal operation of all scientific equipment. These procedures guarantee the reliability and reproducibility of the results obtained.
• Cell Analysis and Imaging Flow Cytometry
  The cell analysis service is available 24/7 for trained researchers, upon reservation through the online booking system. The facility includes advanced technology that allows detailed and complex cellular characterization.
• Cell Sorting
  The cell purification and sorting service is available for booking from Monday to Friday, between 10 AM and 5 PM. Sorting is performed by specialized technicians, ensuring high precision and cell viability. Reservations must be made through the online system.
• Flow Cytometry Training
  The Facility offers high-quality theoretical and practical sessions to all users. The training covers the principles and fundamentals of Flow Cytometry, as well as applications, including cell sorting techniques and data analysis. Courses are tailored to different experience levels, from beginners to advanced users.
• Data Analysis
  We offer specialized support in Flow Cytometry data analysis, including access to a workstation equipped with FlowJo, using advanced algorithms that allow high-dimensional analyses and facilitate the identification of complex cell subpopulations. In addition, we provide personalized support to ensure greater efficiency and accuracy in interpreting results.
• Consulting in Flow Cytometry Experiments
  We provide expert advice at all stages of Flow Cytometry projects, from experimental planning, panel design, and optimization, sample preparation, data analysis, to result presentation. The consulting service is personalized to maximize the quality and impact of research.

Other services:

• Multidimensional Immunophenotyping for the Characterization of Cellular Biomarkers
  This service provides access to next-generation spectral flow cytometry technology for the immune and functional characterization of complex cellular populations. Through multidimensional immunophenotyping, it is possible to analyze cell phenotypes and states with high precision and detail, enabling the identification and validation of relevant cellular biomarkers in contexts such as immune responses, environmental toxicity, and clinical research. Integrated within an advanced research infrastructure, the service combines cutting-edge technology with scientific consultancy to support projects across the entire translational research.;
• Multidimensional Characterization and Identification of Cellular Biomarkers with Integrated Image Analysis
  This service offers detailed and integrated characterization of cellular phenotypes by combining multiparametric analysis of fluorescent markers with high-resolution cellular image data. This approach enables the identification of rare subpopulations, complex functional states, and cellular dynamics that are not detectable using conventional methods. As a result, researchers can gain a comprehensive view of cellular heterogeneity and the biological processes under investigation.
  Main Applications:
  - In-depth analysis of cellular populations in human samples and animal models;
  - Discovery and validation of biomarkers related to immune responses, disease progression, and environmental exposures;
  - Monitoring of dynamic cellular events and cell–cell interactions in physiological and pathological contexts.
  The BD FACSDiscover™ A8 high-end technology provides an unprecedented capacity to analyze up to 80 cellular parameters simultaneously, combining fluorescence with high-complexity image analysis. This unique integration allows for a comprehensive characterization of cell populations at phenotypic, morphological, and functional levels, significantly expanding the potential to identify complex biomarkers and uncover cellular mechanisms with greater precision.
• Multiparametric Functional Cell Analysis
  This service offers a comprehensive approach to functional analysis of cells in a variety of biological contexts. Parameters such as viability, apoptosis, proliferation, cell cycle, metabolism, signaling pathway activation, and cytokine production are assessed using multiparametric methodologies that provide an integrated readout of cellular function. Intracellular cytokine detection by flow cytometry enables profiling of secretion patterns in response to specific stimuli, enhancing the functional assessment and delivering critical insights into immune activity. This detailed characterization helps detect discrete yet meaningful functional changes, contributing to a better understanding of response mechanisms and supporting the identification of potential functional biomarkers in contexts such as immunotoxicity, inflammation, infection, or drug response.
  Main Applications:
  - Study of functional immune responses under baseline and stimulated conditions;
  - Assessment of immunotoxic or pro-inflammatory effects of environmental compounds;
  - Support for the characterization of the functional activity and immunomodulatory effects of immunotherapies, vaccines, or biological agents;
  - Analysis of morphological changes associated with specific functional states using high-resolution cellular imaging.
  Functional analysis is conducted in an integrated and multiparametric manner, allowing the simultaneous measurement of both phenotypic and functional parameters. The inclusion of image-based analysis adds another layer of resolution, enabling visualization of morphological changes linked to cellular states such as apoptosis, activation, or proliferation. This integrated approach enhances the sensitivity and accuracy in detecting subtle functional changes, even in rare or heterogeneous cell populations.
• Development of Customized Panels and Cell Isolation for Advanced Cellular Characterization
  We provide specialized support in the design, development, and optimization of customized panels for high-dimensional flow cytometry, tailored to the specific goals of each project. This service includes strategic marker selection, fluorochrome compatibility assessment, optimization of staining and permeabilization protocols, and scientific-technical guidance throughout the experimental process. Panels are designed to integrate surface, intracellular, and functional markers, enabling comprehensive and high-resolution characterization, even of rare cell populations or in complex biological contexts. Additionally, specific cellular subpopulations can be isolated for subsequent in vitro functional assays, including their response to immunological, therapeutic, or environmental stimuli.
  Main Applications:
  - Custom panel development for studies in immunology, oncology, neuroinflammation, metabolism, or environmental toxicology;
  - Integration of established markers with exploratory targets, including technical validation support;
  - Isolation of specific cell populations for functional assays or downstream analysis;
  - Panel design suitable for translational studies, including both animal models and human samples.
  This service leverages the flexibility of a state-of-the-art flow cytometry platform capable of analyzing up to 80 cellular parameters in a single assay. This capacity enables highly complex and tailored experimental designs, integrating phenotypic, functional, and morphological markers with exceptional resolution. Expert support ensures the robust and efficient development of each panel according to the specific needs of each research project.

Registration and Training

• All new users must register in the Agendo Science system (https://fcm.agendoscience.com/) and request permission before using the equipment for the first time.
• After approval, a mandatory training session is required to allow autonomous use of the analyzer cytometers (FACSCantoII and FACSCalibur).
• For additional support after training, users can request assistance when scheduling on Agendo.

Equipment Use

• Only the user who made the reservation may operate the equipment; booking on behalf of others is prohibited.
• Using equipment without a prior reservation is not allowed and will be monitored by the Facility team.
• All users must follow the maintenance and biosafety guidelines available on the NMS Research intranet.
• The same rules apply to external users. The price list is available on Agendo.
• Cell Sorting (FACSAriaIII): Reservations for cell sorting are made via pre-booking, with detailed information provided for equipment configuration. Confirmation will be sent by email.
• Any equipment malfunctions must be reported to flowcytometry.facility@nms.unl.pt.

Acknowledgments and Publications

• Acknowledgment of the Facility is required in all publications or presentations that involve the use of equipment or services.
• Example of acknowledgment: "Flow cytometry analysis/cell sorting was performed at the Flow Cytometry Facility at NMS."
• If the Facility team contributed significantly to experimental planning or data analysis, co-authorship should be considered.

The Flow Cytometry Facility at Nova Medical School is deeply integrated into national and international scientific networks, promoting knowledge sharing and the development of new skills in the field of Flow Cytometry.

• FLxFlow - Lisbon Flow Cytometry Network
  The Facility’s manager is the founder of this network, which aims to foster collaboration among institutions in Lisbon, promoting the sharing of best practices and resource optimization. Within the FLxFlow framework, the Facility actively participates in organizing international courses and conferences, contributing to the continuous training of researchers and technicians in the field.
• CoLife Alliance
  The Facility is an integral part of CoLife, an alliance established between six life sciences research institutes located in Lisbon and Oeiras. This alliance aims to maximize available resources by sharing infrastructures, services, and scientific knowledge. Collaboration between institutes promotes synergies that drive innovation, shared equipment usage, and the development of joint scientific projects.
• Member of the International Society for Advancement of Cytometry (ISAC)
  The manager is a member of ISAC, one of the leading global organizations dedicated to advancing Flow Cytometry. This affiliation provides access to advanced training programs, international collaboration networks, and the latest innovations in the field.
• Member of the Iberian Society of Cytometry (SIC)
  Participation in the Iberian Society of Cytometry (SIC) reinforces the Facility's commitment to developing Flow Cytometry in the Iberian Peninsula. This society promotes the exchange of technical and scientific knowledge between professionals from Portugal and Spain, encouraging cross-border collaboration.
• CTLS - Core Technologies for Life Sciences