Apoyo a la investigación
CoreLab
CoreLab
The Core Lab of IIS Galicia Sur is a central laboratory that encompasses the facilities and scientific equipment used in basic and clinical research. Its main objective is to standardize the general and common experimental procedures for the various scientific-technological units and platforms of the Institute to provide technical support.
MAIN OBJECTIVES
- Standardize and provide traceability to technical processes to optimize the achievement of high-quality results.
- Reduce working times to provide a rapid response to researchers’ needs.
- Optimize human resources. The reorganization of common workflows helps relieve the workload of infrastructure managers and specialist technicians, thereby improving the efficiency of response to researchers.
- Optimize technological resources by implementing an equipment management system that maximizes their performance and ensures effective experimental planning.
OUTSTANDING SERVICES
- Sample preparation: processing and preservation of biological samples from different origins, extraction and purification of nucleic acids and proteins, histological preparations, sample preparation for flow cytometry and cell separation, among others.
- Development of fundamental molecular techniques: PCR, qPCR, preparation of plates for ddPCR, preparation of NGS libraries, protein quantification, western blot, ELISA, enzymatic assays, cell counting and microscopic visualization, among others.
- Development of basic cellular techniques: primary cell cultures, cell line culture, cell counting, cell cryopreservation, cytotoxicity assays, among others.
- Technical support for clinical studies in the processing, analysis, preservation, and management of biological samples.
Equipment management
The Core Lab is responsible for managing the shared equipment located in the laboratories. To access this equipment, prior registration in the equipment management system “LabArchives Scheduler” is required.
LabArchives Scheduler is an online application that lists all available equipment, aiming to manage their reservations so that all users can better organize their work, ensuring the availability of the necessary equipment for their experimental workflow.
To register, you must complete the registration form and send it to corelab@iisgsgaliciasur.es.
The Cytometry Unit of the IIS Galicia Sur provides technical support and advice, mainly to the IISGS scientific community, but also to the medical community, in the optimization and development of methodologies related to flow cytometry.
Equipment
Cytoflex S Beckman Coulter cytometer.
Technical characteristics:
- 4 Lasers: Violet (405 nm), Blue (488 nm), Green-Yellow (561 nm) and Red (638 nm)
- 15 parameters: 13 Fluorescences (4 violet lasers, 2 blue lasers, 4 green-yellow lasers and 3 red lasers) and forward scatter (FSC) and side scatter (SSc) detectors.
- Side scatter resolution (SSc): 80 nm
- Acquisition speed: 30,000 events/second
Cytoflex SRT Beckman Coulter cell separator.
Technical characteristics:
- 4 lasers: Violet (405 nm), Blue (488 nm), Green-Yellow (561 nm) and Red (638 nm).
- 17 parameters: 15 Fluorescences (5 violet lasers, 2 blue lasers, 5 green-yellow lasers and 3 red lasers) and forward scatter (FSC) and side scatter (SSc) detectors.
- Microplate module.
- Aerosol evacuation system.
- 4 simultaneous separation paths.
- One-way sorting for 6, 24, 48, 96 or 384-well plates.
- Sample acquisition and separation speed: 25,000 events/second.
- Sensitivity for the detection and isolation of particles < 200 nm.
Services
Flow cytometry is an indispensable technique for biomedical research and can be used in all areas of study, both in basic research and clinical research.
Using this technique, the frequency and phenotypic characteristics of immune populations can be determined; assays of cell cycle, proliferation, and cell death and the expression of intra- and extracellular proteins. Furthermore, our cytometer allows the identification and characterization of extracellular vesicles and exosomes.
On the other hand, the cell sorter allows the isolation of specific cell populations, which can be used to carry out -omic techniques (proteomics, genomics, transcriptomics) and functional assays, such as proliferation and viability assays, migration and cell invasion, angiogenesis assays and cell activation assays, among others.
- Responsables: Samuel García Pérez / Beatriz Malvar Fernández
- Email: citometria@iisgaliciasur.es
The Cell Culture Unit of IIS Galicia Sur provides scientific and technical support in animal and human cell cultures while offering different services for diagnostic or basic research purposes, including training of health professionals and the development of new protocols.
Equipment
- Cell culture laboratory equipped to work simultaneously in two laminar flow cabinets for cell culture (Telstar Bio II Advance) with vacuum systems. The room also includes two incubators for cell culture with controlled temperature and CO2 (Memmert INCO 246 med), two centrifuges (Beckman Coulter Allegra X-22R and Eppendorf 5810), an inverted microscope (Leica DMIL LED), a water bath, a -20°C freezer, and a refrigerator.
- BSL-2 cell culture laboratory with negative pressure for handling human samples associated with moderate-risk diseases or for the production of viral vectors for gene therapy. The laboratory is equipped with a laminar flow cabinet (Telstar Bio II Advance), an incubator with humidity and CO2, a freezer, a centrifuge, and a water bath.
- Other infrastructures:
- Real time monitorization of cell growth with ACEA Bio xCELLigence RTCA DP
- Plate reader Telstar Omega (fluorescence, bioluminescence, absorbance).
- Ultracentrifuge Optima XPN-100 (Beckman Coulter).
- Corning Cell counter (Corning).
- Freezer (-80ºC)
- CBS Nitrogen tank (-130 ºC)
- Autoclave Selecta Presoclave II
Services
- Lymphocyte, monocyte and dermal fibroblasts primary culturing
- Maintenance of primary and immortalized cell lines from biological samples (both freezing and proliferation) and mycoplasma detection in cultures (through PCR detection kit)
- Drug cytotoxicity assays (MTT or real time)
- Collection and isolation of primary cultures and transfection of cell lines
- Enzyme activity assays in cell lines
- Real-time monitoring migration and cell adhesion studies
- Viability and cell proliferation assays
- Mononuclear cell isolation from peripheral blood
- Responsables: Saida Ortolano
- Email: cultivos.celulares.iisgs@sergas.es
The Genomics Unit at IIS Galicia Sur provides technical support and guidance for the setup and development of methodologies related to genomic analysis and gene expression studies.
The Unit’s facilities include separate areas for DNA extraction, RNA extraction, pre-PCR, and post-PCR, adhering to the quality standards for such units.
In recent years, its capabilities have been enhanced with the acquisition of an automated digital PCR analysis equipment (in addition to an ultracentrifuge and a bioanalyser) thanks to the grant by the Spanish State Research Agency (AEI) in the call for acquisition of scientific and technological equipment 2018, and the acquisition of a next-generation MiSeq sequencer, which was launched in 2023, using internal funds.
Equipment
- MiSeq Sequencer (Illumina).
- QX200 DROPLET DIGITAL PCR (BioRad).
- CFX Connect Real-Time PCR System (BioRad).
- 7300 Real-Time PCR System (Applied Biosystems).
- QIACube Automated Nucleic Acid Extractor (Qiagen).
- 2100 Bioanalyzer (Agilent).
- Qubit 4 Fluorometer (Life Technologies).
- Biodrop µLite Spectrophotometer.
- ChemiDoc XRS Image Processor (BioRad).
- Sequencher 5.4.6 Genetic Analyzer (Gene Codes).
- Basic Molecular Biology Equipment (thermal cyclers; UV cabinets for pre- and post-PCR; microcentrifuges; centrifuges; horizontal electrophoresis systems; UV transilluminator, etc.).
Featured services
High-Throughput Sequencing – MiSeq
The MiSeq system is a next-generation sequencer using Illumina’s sequencing-by-synthesis technology, known for its accuracy and reliability. This ensures the generation of high-quality, precise data. Additionally, it can generate up to 15 gigabases of sequences per-run, facilitating the analysis of DNA samples of varying sizes and complexities, from small projects to complete microbial genomes. This makes MiSeq a versatile tool for various genomic and clinical research. Moreover, the system comes with user-friendly software that allows experiment design, real-time monitoring, and basic analysis of generated data.
MiSeq Applications:
- Identification of somatic and/or germline variations implicated in various pathologies through the sequencing of specific panels.
- Transcriptomic analysis via RNA sequencing to identify and determine gene expression levels in cells or tissues.
- Microbial genome studies (e.g., SARS-CoV-2, HIV).
- Methylation analysis to identify changes in gene expression activation patterns that may be related to diseases and other biological processes.
- Identification of microorganisms in complex biological samples through 16S rRNA gene sequencing.
- Single-cell studies in collaboration with the Flow Cytometry Unit, using Cell Sorter technology to identify molecular differences between individual cells in a population.
Droplet Digital PCR Analysis – ddPCR
The digital droplet PCR (ddPCR) combines water-oil emulsion droplet technology with microfluidics. Up to 20.000 drops are automatically generated to amplify the sample by PCR in each of them independently. This allows absolute quantitification of target sequences without reference to endogenous patterns or controls. Additionally, it is a highly sensitive, precise, and reproducible technique, especially with low-concentration target sequences, making it ideal for analyzing complex samples (FFPE, miRNA, cfDNA…), detecting rare events (somatic mutations, mosaicism…), and early detection (biomarkers, treatment response, pathogen detection…).
ddPCR Applications:
- Absolute quantification (Viral load, biomarkers, chimerism…).
- Rare event detection (viruses, liquid biopsy, somatic mutations…).
- Gene expression (miRNA and lncRNA, methylation, Single Cell…).
- Copy number variation (CNVs).
- Genomic editing (HDR/NHEJ, DNA rearrangements).
- NGS result validation, library quality, and quantification.
Other Services
- Nucleic acid extraction and QC from various biological samples.
- Quantitative and conventional PCR.
- Sanger Sequencing.
- Genetic diagnosis of neuromuscular diseases: Currently, standardized protocols are available for the study of a wide range of genes involved in hereditary neuromuscular diseases.
- Possibility of optimizing and developing genetic study protocols for other pathologies based on the most appropriate methodological strategies.
- Support in the interpretation of NGS analysis results.
- Training of research and clinical staff through theoretical and practical courses on genomics and molecular biology.
- Responsables: Irene Viéitez
- Email: genomica.iisgs@sergas.es
This unit is designed for the study, detection, and identification of cellular structures in tissue sections and cell cultures through histological and histochemical staining, as well as the use of fluorescent markers. It is further equipped with the Leica 6500 Laser Microdissection system, a microscopic technology that allows for the isolation of small cell groups or even individual cells from heterogeneous tissue or cell cultures for the analysis of DNA, RNA, or proteins from the homogeneous sample obtained.
In addition, the unit has a high-resolution confocal microscope, the Leica Stellaris 8, which offers a central system with a white laser (WLL 440-730 spectrum). This system allows for Fluorescence Lifetime Imaging (FLIM) using TauSense technology, providing immediate access to functional information such as metabolic state, pH, ion concentrations, and more. It eliminates unwanted fluorescence, measures the lifetime of fluorochromes, and enables the separation of overlapping spectra. Moreover, it is equipped with a tool that allows for the direct detection and quantification of molecular interactions using both FLIM and FRET techniques.
It is also worth noting that the system is equipped with an incubation chamber for in vivo cell studies. Additionally, the unit is supported by the necessary software for image analysis.
Equipment
- Tissue processor, embedding station (EG 1500H), microtome (Leica CM 3050), and cryostat (Leica CM 3050)
- Stereomicroscope (M125. Leica)
- Brightfield microscope (DRMBE and DM2000. Leica)
- Fluorescence microscope (DMI6000. Leica)
- Laser microdissector (MLD6500. Leica)
- Super-resolution confocal microscope (STELLARIS 8 Confocal. Leica)
Services
- Sectioning of histological tissue in paraffin and frozen sections
- Brightfield microscopy with photodocumentation
- Fluorescence microscopy with photodocumentation
- Laser microdissection of samples in brightfield or fluorescence
- Development of protocols for sample processing for microscopy (fixation, freezing, staining, immunolabeling, etc.)
- Technical advice for the planning of research projects requiring the use of the laser microdissector
- Responsables: Susana Teijeira / Ainhoa Rodríguez Tébar
- Email: microscopia.iisgs@sergas.es
- Teléfono: 648 912 372
The Biomarkers Platform at IIS Galicia Sur aims to promote and facilitate the development of projects focused on identifying biomarkers of interest in biomedicine. Due to the multidisciplinary nature of these studies, it is essential to adopt a coordinated approach that integrates the various scientific and technological infrastructures within the IIS Galicia Sur research support units. These units have the capacity to identify essential biomarkers for the development of new therapies and personalized treatments.
Key Equipment
- Luminex 200 System Analyzer (Thermo Fisher)
- MiSeq Sequencer (Illumina)
- CytoFLEX SRT Cell Sorter (Beckman Coulter)
- Stellaris 8 Confocal Microscope (Leica)
Main Applications
- Quantification of biomarkers of interest in various pathologies such as pro-inflammatory cytokines, tumor markers, and growth factors, among others.
- Evaluation of gene expression (RNA-seq), analysis of genomic biomarkers, and identification of genetic signatures of interest.
- Detection of biomarkers in circulating DNA.
- Microbiome study and identification of microbial signatures associated with different diseases.
- Research on biomarkers for the development of new therapies and personalized treatments.
- Study of drug-receptor interaction, dose-response efficacy, bioavailability, and cytotoxicity.
- Detection of biomarkers in cellular subpopulations.
- High-resolution imaging of the localization, co-localization, and distribution of biomarkers within cells and tissues.
- Application of advanced in vivo imaging techniques (e.g., FRET, FLIM) for the quantification of secondary messengers in different cellular compartments, studies of protein-protein interactions, and dynamics of protein movements within the cell.
- Responsables:
- Samuel García Pérez – Unidad de Citometría
- Saida Ortolano – Unidad de Cultivos Celulares y Microscopía
- Irene Viéitez – Unidad de Genómica
- Eva Poveda López – Sistema Luminex 200
- Email: biomarcadores@iisgaliciasur.es
