Unit Managers: Susana Teijeira / Ainhoa Rodríguez Tébar
E-mail: microscopia.iisgs@sergas.es
Teléfono: 648 912 372

• Tissue processor, embedding station, microtome and cryostat
• Stereo microscope
• Clear-field microscope
• Fluorescence microscope
• Laser microdissector
• Confocal Microscope – Leica Stellaris 8

Our platform is designed to study, detect and identify cell structures in tissue sections and cell cultures through histological and histochemical stains and fluorescent markers. In addition, we rely on Laser Microdissection Leica 6500, a microscopic technology which allows to distinguish between relevant and non-relevant cells or tissues for DNA/ARN/protein analysis.

The Microscopy Platform has the necessary software to analyse sample images obtained from such equipment.

• Histological sections cut using microtome or cryostat.
• Clear-field microscopy imaging.
• Fluorescence microscopy imaging.
• Laser microdissection with samples in clear field or with fluorescence.
• Sample processing for microscopy (fixation, freezing, staining, immunolabeling, etc.)
• Technical advice for research projects that require laser capture microdissection.

Unit Manager: Irene Viéitez
E-mail: genomica.iisgs@sergas.es

The Genomics Unit of the Galicia Sur Health Research Institute offers technical support and advice in the development of methodologies related to genetic analysis and gene expression studies helping both the scientific community of the Institute and the medical community. Today, knowledge on genetic bases behind hereditary diseases as well as genetic diagnosis in patients with known pathologies are essential in the application of treatments and the development of new therapies specific to the patient’s genotype.

The Genomics Unit has infrastructure and technological resources in the research laboratories of the Álvaro Cunqueiro Hospital. The Unit is supported by the DNA Sequencing Service from the CACTI at the University of Vigo.

Facilities of the Genomics Unit are located on the second floor within the technical block, and include independent areas for DNA, ARN, PCR and post-PCR extraction in compliance with the quality standards of these type of units.

• QX200 DROPLET DIGITAL PCR (Biorad)
• ABI PRISM 310 Genetic Analyzer (Applied Biosystems)
• 2720 Thermal Cycler (Applied Biosystems)
• GeneAmp PCR System 9700 thermal cycler (Applied Biosystems)
• Biodrop µLite Spectrophotometer
• 7300 Real Time PCR System (Applied Biosystems)
• Eppendorf Thermomixer C (Eppendorf)
• ChemiDoc XRS System (BioRad) / ImageQuant350 (GE Healthcare)
• Microcentrifuges: Mikro200 (Hettich) and Allegra X-22R (Beckman Coulter) Horizontal electrophoresis systems
• Molecular Biology basic equipment (refrigerators; freezers; deepfreezers; thermostatic bath; centrifuges; UV transilluminator; Milli-Q Water Purifier System…)

The IISGS Genomics Unit has recently strengthened its infrastructures after 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.

Digital PCR technology is based on partitioning a sample into many individual PCR reactions in such a way that some of these partitions contain the target sequence (positive) while others do not (negative). After PCR, the fraction of negative partitions is used to obtain absolute and accurate quantification of target sequences without reference to endogenous patterns or controls.

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 quantities to be measured by counting molecules in discrete and volumetrically defined droplets. The reading of the fluorescently positive and negative drops enables calculation of the concentration of DNA.

Our services are aimed at both basic research as well as genetic diagnosis, and range from advice on experimental design to the issue of results.

General Services:

Obtaining nucleic acids from different biological samples:

• Extraction
• Purification
• Quantification

Conventional PCR:

• Oligonucleotide design and optimisation of PCR conditions
• Amplification by simple, multiple or nested PCR
• PCR-RFLP analysis with restriction enzymes
• Interpretation of agarose gel electrophoresis results

DNA sequencing:

• Analysis of mutations
• Resequencing of variants
• Genotyping (SNP analysis)
• Fragment analysis
• Sequence reading and interpretation

Real-Time PCR:

• Oligonucleotide design and Real-time PCR protocol optimization
• Relative or absolute quantification studies in gene expression
• Allelic discrimination studies using TaqMan probes
• Genomic copy number variation determination
• Result analysis

Specific Services:

• Genetic diagnosis of neuromuscular diseases: We currently have standardized protocols for the study of a wide variety of genes involved in hereditary neuromuscular diseases.
• Development and optimization of genetic study protocols for diseases based on the most suitable methodological strategies.
• Support in the interpretation of exome analysis results.
• Training of researchers and health staff through theoretical and practical courses on genetics and molecular biology.

Our services are aimed at both basic research as well as genetic diagnosis, and range from advice on experimental design to the issue of results.

General Services:

Obtaining nucleic acids from different biological samples:

• Extraction
• Purification
• Quantification

Conventional PCR:

• Oligonucleotide design and optimisation of PCR conditions
• Amplification by simple, multiple or nested PCR
• PCR-RFLP analysis with restriction enzymes
• Interpretation of agarose gel electrophoresis results

DNA sequencing:

• Analysis of mutations
• Resequencing of variants
• Genotyping (SNP analysis)
• Fragment analysis
• Sequence reading and interpretation

Real-Time PCR:

• Oligonucleotide design and Real-time PCR protocol optimization
• Relative or absolute quantification studies in gene expression
• Allelic discrimination studies using TaqMan probes
• Genomic copy number variation determination
• Result analysis

Specific Services:

• Genetic diagnosis of neuromuscular diseases: We currently have standardized protocols for the study of a wide variety of genes involved in hereditary neuromuscular diseases.
• Development and optimization of genetic study protocols for diseases based on the most suitable methodological strategies.
• Support in the interpretation of exome analysis results.
• Training of researchers and health staff through theoretical and practical courses on genetics and molecular biology.

Unit Manager: Saida Ortolano
E-mail: cultivos.celulares.iisgs@sergas.es

• Autoclave Selecta Presoclave II
• Water bath J.P. Selecta Precisdig 6001187
• Two Class II Biological Safety Cabinets Telstar Bio II Advance
• Eppendorf 5810 Centrifuge + accessories
• Beckman Coulter Allegra X-22R Refrigerated Centrifuge + accessories
• Radiber CVE-480-25 Vertical Freezer (-20°C)
• Two Thermo Scientific Vertical Freezers (-80°C)
• Automated Cell Coubter Millipore Scepter 2.0
• Air Liquide and MVE Lab Series liquid nitrogen dewars
• ACEA xCELLigence Real Time Cell Analysis (RTCA) Instrument
• Twinkle LB 970 Microplate Fluorometer – Berthold Technologies
• Radiber FFD 500 Refrigerator
• Two Memmert CO2 Incubators INCO246Med
• Biochrom Asys UVM340 Microplate Reader
• Leica DM IL LED Inverted Microscope

IISGS has two culture rooms. One of them is dedicated to working with Adeno-Associated Virus transfected cells.

In addition, a P2 Laboratory (BSL-2) has been recently installed, equipped with biological safety cabinets, negative pressure system, airlock entry and other physical containment equipment, making it suitable for working with moderately hazardous agents associated with human diseases (i.e. pathogenic or infectious organisms).

The Cell Culture Unit 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.

Services can be performed by the Unit technician or on a self-serve basis. Our techniques range from basic services to the study and characterization at the molecular and cellular level. Our techniques include:

• 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

Unit managers:  Samuel García Pérez / Beatriz Malvar Fernández
E-mail: citometria@iisgaliciasur.es

La citometría de flujo es una técnica indispensable para la investigación biomédica y puede usarse en todas las áreas de estudio, desde la investigación más básica hasta las aplicaciones más clínicas.

La Unidad de Citometría del IIS Galicia Sur da soporte técnico y asesoramiento, principalmente a la comunidad científica del IISGS, pero también a la comunidad médica, en la puesta a punto y desarrollo de metodologías relacionadas con la citometría de flujo.