The High-standard Technological services (STANs) comprise activities such as assays, technical assistance and training, for which the equipment, infrastructure and human resources are granted by INIBIOLP. STANs are offered to several areas of society.
STAN Specifications: Analysis of fatty acids by gas chromatography.
Methodology: Analysis by CGC: An Omega Wax 250 column of 30 m will be used.
The operative conditions will be as follows: injector in split mode at 260 260°C, programmed oven temperature, and flame ionization detector will be maintained at 280°C.
Helium will be the gas carrier at a flux of 1-2 ml/min.
Equipment: Gaseous Chromatograph Hewlett Packard 6890.
STAN Specifications: Analysis by gaseous chromatography – mass spectrometry for the detection / identification of compounds or organic contaminants.
Methodology: 1) Determination of optimal operational conditions of sample nature. Sample taking (liquid or gaseous) and injection. Deliver of chromatogram and spectra of interest in printed or electronic version.
2) Identity assignment by comparison of spectra obtained with spectra commercial libraries (NIST 05/EPA/NIH, pesticides, essential oils and others) and with other spectra obtained from standards (provided by the applicant).
Equipment: Capillar gas chromatograph with spectrometry detector of mass Agilent Technologies CGC-HP6890 MS5975VL.
STAN Specifications: Fractioning of biological samples by differential ultracentrifugation.
Methodology: The fractioning of biological samples will be performed previously to adequate rotor selection and rotation velocity, solvent density, etc.
Installation startup, sample loading, and ultracentrifugation.
Equipment: Beckman Coulter LE-80K Preparative Ultracentrifuge.
STAN Specifications: Evaluation of insecticide capacity. Analysis of insecticide efficacy of biological, physical or chemical insecticide agents by means of mortality determination, effect on life cycle of insects, etc, and/or evaluation of lethal doses.
Methodology: Bioassays will be performed by exposing insects to compounds to be evaluated. The application mode depends on the physicochemical properties of the compound and/or as it was solicited. Batches of at least 10 insects will be employed, with at least 4 replicas for each concentration or condition to be assayed. Statistical analysis of the results.
STAN Specifications: the service is focused on purifying or separating biomolecules as proteins, peptides, polynucleotides, among others, through the use of liquid chromatography (FPLC or HPLC) employing columns of anionic exchange..
Methodology: It is used an anionic exchanger column (Mono-Q 10/100 GL) employing gradients of solvents of growing ion force. The detection is performed with a UV-Vis detector. The service has the possibility of thermostatizing the sample between 4 and 40 °C.
Tubes are collected and submitted at the end of the STAN together with a report of the results, such as chromatograms and tables for the applicant’s later concentration and analysis. No certification will be granted for the activities involved in the present STAN.
Equipment: Chromatographs HPLC Agilent Technologies 1260 Infinity model and HPLC Waters ARC model.
Service Manager: Qca. Letizia Bauzá Scientific Supervisor: Dr. Gisela Franchini
STAN Specifications: the service is focused on identifying, quantifying and/or separating analytes, through the use of HPLC employing reverse phase columns with different resolution properties.
Methodology: Reverse phase columns are used (C18, C8, etc.). We also offer the possibility of thermostatizing the sample between 4 and 40 °C and the column between 4 and 60 °C, using isocratic methods or in solvent gradients and UV-Vis detection.
For IDENTIFICATION and/or QUANTIFICATION: it is done a calibration curve with the standard (each point and each injection in duplicate).
PURIFICATION and/or SEPARATION: it is performed the tube collection which are submitted for the applicant’s later concentration.
A report with the results such as chromatograms and tables is also submitted for the applicant’s later analysis.
No certification will be granted for the activities included in the present STAN.
Equipment: HPLC Waters ARC model.
Service Manager: Qca. Letizia Bauzá Scientific Supervisor: Dr. Gisela Franchini
STAN Specifications: this service is tailored to the purifications of proteins and other macromolecules through liquid chromatography (FPLC or HPLC) using molecular exclusion. Columns with different ranges of size are utilized.
Methodology: Columns of molecular exclusion are used to separate the following macromolecules.
Proteins in the following ranges:
* 3-70 KDa (Superdex 75 10/300 GL)
* 100-600.KDa (Superdex 200 10/300 GL)
* 5-5.000 KDa (Superosa 6 30/100GL)
Dextrans in the following ranges:
* 500 -30.000 Da (Superdex 75 10/300 GL)
* 1-100 KDa (Superdex 200 10/300 GL)
* 1-300 KDa (Superose 6 30/100GL).
The service has the possibility of thermostatizing the sample between 4 and 4°C. Analytes with aqueous buffer are eluted (TRIS, PBS) in an isocratic elusion. Detection is carried out measuring the absorbance with detector UV-VIS
For purification and/or separation, the applicant must ensure that the samples are usable for the chromatographic analysis (filtered and/or centrifuged previously). The maximum volume that is injected for the run is of 500 microliters (it is considered one sample). Tubes are collected and submitted to the applicant for its later concentration. The samples can be alike or different provided that the same chromatographic conditions are maintained.
Equipment: HPLC Waters ARC model.
Service Manager: Qca. Letizia Bauzá Scientific Supervisor: Dr. Gisela Franchini
STAN Specifications: Determination of drug or compound toxicity in cell culture
Methodology: The basic service includes the cell incubation with the drug o compound of interest, its staining with crystal violet or the determination of the mitochondrial activity by standard colorimetric methods (3-(4,5-dimetylthiazol)-2,5-diphenyltetrazolium (MTT) bromide) in order to evaluate the effect of the drug/compound on the cell proliferation/viability. Both the cell line and the experimental design to be used will be coordinated with the user based on the availability of the institute and on the requirements of the study to be carried out.
The statistical analysis of the results, the LD50 determination and other more specific analysis are not included in the basic service, but they can be coordinate with the user.
Equipment: BSC Esco class II Laminar Flow BSC Model ACZ-451, Panasonic CO2 Laboratory Incubator Model MCO-18AC-PE, Beckman Coulter DTX 880 Multimode Detector Plate Reader.
STAN Specifications: Real-time quantitative PCR analysis.
Methodology: The service includes the relative quantification of genes of interest, normalized with reference genes in problem samples compared to control samples. It can include, on the request of the interested party, the previous nucleic acid extraction (RNA, DNA), the cDNA synthesis, the construction of calibration curves, and the data statistical analysis.
Equipment: Real-Time PCR Agilent Technologies Aria Mx
STAN Specifications: System for the quantification of equivalent genomes of virus SARS-COV-2 in naso-pharyngeal swab samples and other biological samples (RUO).
Methodology: It consists of a one-step qRT-PCR (R.U.O.) assay that allows the accurate detection and quantification of SARS-CoV-2 viral load through the analysis of two viral genes (ORF1ab and E gene) and a reference gene (RNAseP) on viral RNA samples obtained from several biological samples. The samples may consist of naso-pharyngeal swab tests, cerebrospinal fluid (CSF), saliva, fecal matter, blood or any other human or animal biological sample, or of inert material such as raw sewage, where virus SARS-CoV-2 can be detected. These biological samples will be provided by the contracting party according to their study design. All the material will be handled according to the Biosecurity Compliance Requirements. The contracting party will receive a report with the results to be used as epidemiologic diagnosis and/or analysis in accordance with the purpose of the sample derivation.
STAN Specifications: The basic service consists of collecting data from biological samples using the flow cytometer. The samples can be fresh or fixed in tubes, eppendorf tubes or multiwall plaques. Functional studies can also be performed; namely, to monitor the cell response over time against a stimulus. Besides, the processing and marking of biological samples can be sought in order to acquire and/or carry out the analysis of the obtained data.
Certifications can be issued (operational qualification. OQ). Please make the request and they will be done on demand.
Equipment: BD Accuri C6 Plus Flow Cytometer: it has two lasers, blue (488 nm) and red (630 nm). 6 parameters can be measured: FSC, SSC and 4 fluorescence channels (510nm-800 nm).
Technical Supervisor: Julia Tau, support personnel, CONICET.
STAN Specifications: Optimization of culture conditions for the growth and differentiation of several types of fungal propagules (conidia, blastopores, mycelial pellets, microsclerotia) to be used as biosupplies in the handle of plague insects, from strains of fungi provided by the contracting party. Planning and monitoring of biochemical and molecular markers useful as parameters of virulence. Evaluation of the advantages and disadvantages of the different types of propagules.
Methodology: Practical consulting (face-to-face workshops) at the requested laboratory and theoretical consulting with talks (face-to-face or virtual) will be provided. Consulting is tailored to both personnel of public institutions such as universities and laboratories from state institutions and private companies interested in the development and production of bioinsecticides products.
STAN Specifications: Quantification and analysis of samples of nucleic acids through nano-spectrophotometry followed by electrophoresis
Methodology: The service consists of two stages which can be requested together or separately:
1) Precise quantification of DNA, RNA, from 1-2 µL of the sample with a nano-spectrophotometer and analysis of their quality parameters through the relation between the absorvances 260nm/280nm and 260nm/230nm;
2) Visualization, quality control and product detection of DNA, RNA through agarose gels electrophoresis. Gel visualization is performed by a UV transilluminator.
Equipment: Nanodrop Thermo SCIENTIFIC, 2000c Spectophotometer; electrophoresis cube Amersham Biosciences, HE33 mini horizontal submarine unit.
STAN Specifications: Tailored to public as well as private organisms requiring the offered service.
Methodology: Lyophilization consists of the concentration of liquids by sublimation, from a solid state to a water vapor state. This is achieved by freezing at temperatures lower than 50 C and very low pressures. This method allows concentrating or drying samples preserving its properties. The samples are frozen at -80C in suitable containers for vaccum. They are exposed to the lyophilizer for a time period depending on the volume and required conditions.
Equipment: Lyophilizer VirTis 10-030 with manifold valves for bottle-like containers.
STAN Specifications: To advise on bioinformatic and biochemical techniques aimed at molecular and cellular biology.
Methodology: To give advice on methodologies of programming and automatic learning, and also on the biochemical analysis oriented to applications in molecular and cellular biology of eukaryotic and prokaryotic cells.
STAN Specifications: We offer the preparation of lipid vesicles (liposomes) of different composition and size.
Methodology: Liposomes will be prepared with different methodologies depending on their size: liposomes of 50 nm, named Small unillamelar vesicles (SUVs) of 100 or 200 nm by sonication, and large unillamelar vesicles (LUVs) by extrusion. They can be prepared of different lipid composition and will be charged with fluorophores, ATP, antibiotics, and so on, depending on the client’s need. Lipids and intravesical content will be provided by the client. It will be prepared 1 ml of solution of the requested lipid concentration. Each preparation of liposomes will be accompanied by a report of the actual distribution of the size of the prepared vesicles and their potential of a charge determined by Dynamic light scattering (DLS).
Service Manager: Dr. Sabina Maté and Dr. Vanesa Herlax
STAN Specifications: The acquisition of images and films in a spectral confocal microscopy. Biological samples (cells, tissues, small organisms, nano/microstructured materials, organic molecules). Image acquisition: 2D, 3D, time-lapse, emission/excitation spectrum, line scanning, life lapses, STED techniques, FRAP, FLIP, FRET, FLIM-FRET. It is directed to professors, researchers and professionals of public organisms and companies.
Methodology: The confocal microscope combines capacities to implement a great variety of techniques and can be optimized for the detection of a wider variety of fluorescent probes. The use of a white light laser and a 405 nm laser enables to select the best wave length to excite each of the fluorophores, and even perform an excitation spectrum between 440 and 790 nm. Likewise, there are four hybrid cutting-edge detectors optimized for the emission detection between 410 and 850 nm, also enabling the registry of emission spectra. Since the light sources are pulsed and the detectors have ultrarapid electronics, the measurements can be performed in the FLIM mode (fluorescence-timelife imaging microscopy). Also, it has a STED laser of 775 nm to obtain superresolution images (between 30 and 70 nm) for fluophores specific of the red and NIR range. And the combination with the technique of life-times enables its exclusive application in live samples (TauSTED). The microscope has accessories to work with live cells and small organisms such as Caenorabdithis elegans (C.elegans, a free living nematode). The service does not include an assay report, data and obtained images will be delivered.
Equipment: Leica Stellaris 8 FALCON STED (Stimulated Emission Depletion) Microscope, Leica Microsystems.
