IDIBAPS, Institut d'Investigacions Biomèdiques August Pi i Sunyer

Services:

Transmission electron microscopy (TEM)

• Ultrastructure studies of tissues and cell cultures (on different subtrates)
• Studies of suspensions (bacteria, virus, flagella, lipid vesicles...)
• Studies of cellular organelles extraction (reticulum, mitochondria, exosomes...)
• Molecular localization and colocalization of proteins, glucidic residues,..
• Molecular localization of DNA or RNA sequences by “in situ” hybridation
• Cytochemical studies (enzymatic activity, enzymatic digestions..)
• Correlative techniques (optical, confocal and electron microscope)
• Preparing of samples for TEM-based microanalysis
• Studies of viruses, organules and proteins by negative staining.

Scanning electron microscopy (SEM)

• Microstructural analysis of surface cell cultures on different substrates (glass, thermanox, aclar,..)
• Surface characterization of tissues or organs
• Studies of cellular suspensions(bacteira...)
• Studies of surfaces: bony, teeth, cornea, skin,...
• Characterization of biofilms and their formation
• Morphological studies of small organisms and micro-organisms
• Surface molecular localization
• Characterization of biomaterials, implants...
• Study, analysis and assessment of the images

Examples of applications

• Biomedical characterization of any kind of cell culture or tissues by SEM or TEM
• Studies of markers incorporation (colloidal gold, quantum-dots..) inside the tissues or cell culture (ex. Inhalation of nanogold and their localisation by light and transmissionn electron microscopy)
• Sperm cell study by TEM and SEM for diagnosis
• Studies of biofilms on water filters, tracheal tubes, historical monuments.
• Characterization of protein aggregates for example in Alzheimer’s disease
• Biopsies in pathology
• Calcium microanalysis detection

Sample preparation types:

Cryomethods preparation (cryofixation)

• High pressure cryofixation, cryosubstitution, cryo embedding in epoxy resins ( for ultrastructural studies)
• High pressure cryofixation, cryosubstitution, cryo embedding in acrylic resins at low temperatures (molecular location studies: immunolocalization, "in situ" hybridization, etc.)
• High pressure cryofixation, cryosubstitution, critical point process (3-D studies using scanning electron microscopy)

Standard preparation method (chemical fixation)

• Chemical fixation, dehydration and embedding in epoxy resins for ultrastructural studies
• Ultramicrotomy at room temperature
• Negative staining (viruses, flagella, proteins, etc.)
• Cytochemical techniques (detection of enzymes, glycoproteins, sugars, etc.)
• Chemical fixation, dehydration, critical point (scanning electron microscopy)

Mixed preparation: combination of standard methods and cryomethods

• Chemical fixation, cryoprotection, cryosubstitution and embedding in epoxy resins (ultrastructural study)
• Chemical fixation, cryofixation and embedding in epoxy resins (ultrastructural study)
• Chemical fixation, cryoprotection, cryosubstitution and embedding at low temperatures (molecular localization studies: immunolocalization, "in situ" hybridization, etc.)
• Chemical fixation, cryoprotection, cryofixation and cryoultramicrotomy (molecular location studies: immunolocalization, "in situ" hybridization, etc.)
• Chemical fixation, cryofixation, cryosubstitution, critical point process (for scanning electron microscopy)

Molecular localization

• Proteins, sugar residues,...
• Markers: nanogold, silver enhancement, colloidal gold, quantum-dots, lectins.

Correlative techniques

• To observe the “same” cell (for example a dynamic process) by light microscopy (at low resolution) and by transmission electron microscopy (higher resolution) using cryofixtion or chemical fixation.

Equipment:

• Jeol EM 1010 transmission electronic microscope
• Zeiss DSM 940A scanning electron microscope
• EM PACT cryofixation system (Leica)
• Cryosubstitution systems
  • AFS (Leica)
  • AFS2 (Leica)
• Ultramicrotomes
  • Ultracut (Reichert-Jung)
  • Ultracut E (Reichert-Jung)
  • Ultracut UCT (Leica) + FCS Cryoultramicrotomy Unit (Leica)
• VGMicrotech CPD 7501 critical point system
• Fisons Instrument FC510 Sputtering System
• Optical microscopes
  • Leica DMLS
  • Olympus CX41 RF + Olympus DP 70 digital camera