Facilities and equipment
The group has state-of-the-art technology to conduct cutting-edge research in reproductive biotechnology.
Facilities
Our laboratories at the University of Girona, where we carry out our day-to-day research.
Equipment
CASA (Computer-Assisted Sperm Analysis)
Automatic system for analysis of sperm movement using computer vision. Measures kinematic parameters of motility, average velocity (VAP), displacement velocity (VSL), curvilinear velocity (VCL) and linearity index (ALH) according to standardized criteria for semen quality assessment.
CytoFLEX flow cytometer
Multiparametric equipment for high-throughput cell population analysis, equipped with 3 lasers (405, 488 and 638 nm) and 10 emission filters. Allows simultaneous evaluation of multiple functional markers in spermatozoa: viability, acrosomal integrity, mitochondrial membrane potential, ROS generation, apoptosis, DNA fragmentation and de novo RNA synthesis.
Fluorescence microscope
Direct and inverted epifluorescence systems equipped with high-magnification objectives and optical setups for visualization of tissues and cells labeled with fluorophores. Applied to analysis of viability markers, acrosomal integrity, chromosomal damage and intracellular protein localization using immunofluorescence.
Programmable Rate Freezer
Controlled freezing system for cryopreservation of gametes and embryos. Allows establishment of precise and reproducible cooling rates, optimizing ice crystal formation and minimizing damage from osmotic stress during the freezing process.
Fluorimeter
Fluorescence spectrophotometer for measuring fluorescence emitted by biomolecules in solution. Applied to evaluation of plasma membrane integrity, mitochondrial activity and other sperm quality markers using fluorescent probes.
Spectrophotometer
Instrument for measuring absorbance and transmittance of solutions at specific wavelengths. Allows assessment of protein concentration and purity, nucleic acids and other biomolecules, as well as colorimetric assays based on enzymatic reactions.
Real-time and conventional thermocycler
Equipment for Polymerase Chain Reaction (PCR). Conventional thermocycler performs nucleic acid amplification with endpoint detection; real-time thermocycler (qPCR) allows continuous monitoring of amplification during each cycle, enabling relative and absolute quantification of mRNA and DNA.
Laminar flow hoods
Sterile work spaces equipped with HEPA filtration that maintain an environment free of contaminant particles. Essential for cell culture techniques, sample preparation and manipulations requiring sterility.
Timelapse chambers for embryos (CytoSMART LUX2)
Incubation system with automatic image capture at regular intervals without removing embryos from the incubator. Allows continuous monitoring of embryonic development, morphokinetic analysis and identification of implantation predictive biomarkers based on division timings.
Liquid nitrogen tanks
Cryopreservation systems for long-term storage of semen, oocytes and embryos at -196 °C. Nitrogen tanks, including dry nitrogen systems, maintain temperature stability and allow indefinite preservation of gametes and reproductive material.
Micromanipulator
Precise micromanipulation equipment for intracytoplasmic sperm injection (ICSI), oocyte aspiration and embryo manipulation. Allows three-dimensional movements of micrometric precision controlled manually or motorized.
Take3
Instrument for measuring concentration and purity of proteins and nucleic acids in ultra-reduced volumes (1-2 μL). Allows rapid assessment of protein content, DNA/RNA integrity and degree of contamination in laboratory samples without requiring cuvettes.
TissueLyser
Tissue and cell homogenizer using high-speed bead beating for the mechanical disruption of biological samples. Enables efficient and reproducible extraction of nucleic acids and proteins from tissues, processing multiple samples simultaneously.
Osmometer
Instrument for measuring the osmolality of solutions and media by freezing-point depression. Essential for quality control of semen extenders, cryopreservation media and assisted reproduction solutions, ensuring optimal osmotic conditions for the survival of gametes and embryos.
Basic laboratory equipment
Quality-filtered gas hood, autoclave for steam sterilization, pH meter for pH control in culture media, tube and microtiter plate centrifuges, cold chambers (4 °C) for storage of heat-sensitive reagents, -20 °C freezers for sample preservation and -80 °C freezers for long-term storage of samples and biological reagents.
SensorDish® Reader (PreSens GmbH)
24-well plate reader equipped with optical fluorescence sensors for real-time measurement of dissolved oxygen consumption in culture media. Allows simultaneous evaluation of multiple samples with oxygen concentration recordings at 30-second intervals, under controlled temperature (38 °C) and atmosphere (5% CO₂) conditions, essential for the study of mitochondrial respiration and the energy metabolism of reproductive cells.
Techniques
Seminogram (CASA)
Evaluation of seminal parameters —concentration, motility, kinetics and morphology— using CASA systems (Computer-Assisted Sperm Analysis). Allows objective and standardized analysis of sperm quality according to WHO criteria.
Sperm cryopreservation
Freezing and vitrification protocols optimized for long-term preservation of semen from animal and human species. Includes assessment of cryotolerance and identification of predictive biomarkers for post-thaw survival.
In vitro fertilization
In vitro oocyte maturation (IVM), sperm capacitation, fertilization and embryo culture procedures. Application in animal models and assisted reproduction technologies.
Flow cytometry
Multiparametric analysis of sperm populations at high speed. Simultaneous evaluation of about ten functional markers, including: viability (SYBR-14/PI), acrosomal integrity (PNA-FITC), mitochondrial membrane potential (JC-1), reactive oxygen species (CellROX, H2DCF, HE, MitoSOX), apoptosis (Annexin V/PI), DNA fragmentation (TUNEL), and other cellular damage markers.
Comet assay
Single cell gel electrophoresis (SCGE) for detection of single-strand (alkaline assay) and double-strand (neutral assay) DNA breaks. Requires complete decondensation of sperm chromatin for proper analysis.
TUNEL
Enzymatic labeling of fragmented DNA 3'-OH ends using terminal deoxynucleotidyl transferase (TdT). Assessable by both fluorescence microscopy and flow cytometry. Direct and sensitive method for quantifying sperm DNA fragmentation.
SCSA (Sperm Chromatin Structure Assay)
Evaluation of sperm chromatin structure using acridine orange and flow cytometry. Provides DNA fragmentation index (DFI) and fraction of spermatozoa with immature chromatin (HDS).
SCD (Sperm Chromatin Dispersion)
Chromatin dispersion test based on acid denaturation and removal of nuclear proteins. Generates characteristic dispersion halos allowing classification of spermatozoa according to DNA integrity. Available in alkaline and neutral variants.
DNA quantification by fluorimetry
Accurate measurement of DNA concentration using a fluorometer and fluorophores that selectively bind double-stranded DNA. Provides more sensitive and specific quantification than spectrophotometry, especially in low-concentration or contaminant-containing samples.
Western blot
Detection and semi-quantification of specific proteins in cell extracts using polyacrylamide gel electrophoresis (SDS-PAGE), membrane transfer and immunodetection with antibodies.
Immunofluorescence
Visualization of subcellular localization of specific proteins using antibodies conjugated with fluorophores. Applied to study proteins involved in capacitation, acrosome reaction and sperm cell signaling.
Confocal microscopy
Obtaining images of tissues and cells with high spatial resolution in three dimensions. Allows colocalization of multiple fluorescent markers and three-dimensional reconstruction of cellular compartments.
Electron microscopy
Ultrastructural observation of spermatozoa and reproductive tissues at high resolution. Includes transmission electron microscopy (TEM) and scanning electron microscopy (SEM) for detailed morphological analysis.
Cell cultures
Maintenance and growth of cells under controlled conditions. Application in in vitro models for studying reproductive physiology, toxicology and functional assays.
RT-qPCR (Real-time PCR)
Relative or absolute detection and quantification of mRNA and other nucleic acids in real time. Used for analysis of gene expression in spermatozoa, testicular tissue and reproductive cells.
Proteomics
Massive identification and quantification of proteins using mass spectrometry (LC-MS/MS, TMT). Applied to study of sperm proteome, seminal plasma and extracellular vesicles for identification of fertility biomarkers.
Metabolomics
Metabolic profile of sperm cells and seminal plasma using nuclear magnetic resonance (¹H-NMR) and gas chromatography-mass spectrometry (GC-MS). Allows identification of metabolites associated with sperm quality and fertility.
Evaluation of reactive oxygen species (ROS)
Determination of intracellular ROS levels using fluorescent probes (CellROX Deep Red, CellROX Green, DHE) and flow cytometry or fluorescence microscopy. Discrimination between physiological levels and oxidative stress.
Chromomycin A3 (CMA3) test
Evaluation of the degree of protamination of sperm chromatin. CMA3 competes with protamines for binding to minor grooves of GC-rich DNA; elevated fluorescence indicates deficient histone-protamine substitution.
Evaluation of oxygen consumption (OCR)
Determination of mitochondrial respiratory activity of spermatozoa. Allows assessment of metabolic capacity through measurement of oxygen consumption rate.